Inhibitors of sarm1

ABSTRACT

The present disclosure provides compounds and methods useful for inhibiting SARM1 and/or treating and/or preventing axonal degeneration.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/899,444, filed Sep. 12, 2019, which is herein incorporated byreference in its entirety.

SEQUENCE LISTING

The instant application contains a Sequence Listing, which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. The ASCII copy, created Sep. 9, 2020 is named2012800-0041_SL.txt, and is 8,858 bytes in size.

BACKGROUND

Axonal degeneration is a hallmark of several neurological disordersincluding peripheral neuropathy, traumatic brain injury, andneurodegenerative diseases (Gerdts et al., SARM1 activation triggersaxon degeneration locally via NAD(+) destruction. Science 348 2016, pp.453-457, hereby incorporated by reference in its entirety).Neurodegenerative diseases and injuries are devastating to both patientsand caregivers. Costs associated with these diseases currently exceedseveral hundred billion dollars annually in the Unites States alone.Since the incidence of many of these diseases and disorders increaseswith age, their incidence is rapidly increasing as demographics change.

SUMMARY

The present disclosure provides technologies useful, among other things,for treating and/or preventing neurodegeneration (e.g., for reducingaxonal degeneration). In some embodiments, provided technologies inhibitSARM1.

In some embodiments, the present disclosure provides certain compoundsand/or compositions that are useful in medicine, and particularly fortreating neurodegeneration (e.g., for reducing axonal degeneration).

In some embodiments, the present disclosure provides compounds having astructure as set forth in Formula I.

or a pharmaceutically acceptable salt thereof, wherein:

-   X is N or C—R^(x);-   R^(x) is selected from the group consisting of hydrogen, halogen,    and optionally substituted C₁₋₆ aliphatic;-   R^(a) is selected from the group consisting of hydrogen, halogen,    CN, CF₃, C₁₋₃ aliphatic, and —C(O)N(R^(a)*)₂;-   R^(a)* is selected from hydrogen and C₁₋₃ aliphatic;-   R¹ is selected from the group consisting of hydrogen, halogen,    N(R)₂, OR, C₁₋₆ aliphatic, a 3- to 7-membered monocyclic    heterocyclic ring having 1-2 heteroatoms independently selected from    nitrogen, oxygen, and sulfur, a 5- to 6-membered heteroaryl ring    having 1-3 heteroatoms independently selected from nitrogen, oxygen,    and sulfur, and a 7- to 8-membered bridged bicyclic heterocyclic    ring having 1-2 heteroatoms independently selected from nitrogen,    oxygen, and sulfur, wherein each of C₁₋₆ aliphatic, monocyclic or    bridged bicyclic heterocyclic ring, and heteroaryl ring is    optionally substituted with 1-3 R¹* groups;-   R¹* is selected from oxo, halogen, N(R)₂, OR, C(O)R, S(O)₂R, C(O)OR,    C(O)N(R)₂, optionally substituted C₁₋₆ aliphatic, and an optionally    substituted 5- to 6-membered heteroaryl ring having 1-3 heteroatoms    independently selected from nitrogen, oxygen, and sulfur;-   R is selected from the group consisting of hydrogen, an optionally    substituted C₁₋₆ aliphatic, and an optionally substituted 3- to    7-membered saturated monocyclic ring having 0-2 heteroatoms    independently selected from nitrogen, oxygen, and sulfur, or:    -   two R groups, together with the nitrogen atom to which they are        attached, form an optionally substituted 3- to 6-membered        heterocyclic ring containing 0-1 additional heteroatom selected        from nitrogen, oxygen, or sulfur;-   R² is selected from the group consisting of hydrogen, halogen,    S(O)₂R, C(O)OR, C(O)N(R)₂, and an optionally substituted group    selected from C₁₋₆ aliphatic, a 3- to 7-membered heterocyclic ring    having 1-2 heteroatoms independently selected from nitrogen, oxygen,    and sulfur, and a 5- to 6-membered heteroaryl ring having 1-3    heteroatoms independently selected from nitrogen, oxygen, and    sulfur; and-   R³ is selected from the group consisting of hydrogen, halogen, CN,    CF₃, C₁₋₃ aliphatic, and —C(O)N(R^(a)*)₂.

In some embodiments, provided compounds have structures of Formulae I-a,I-a-i, I-a-ii, I-b, I-b-i, I-b-ii, I-c, I-c-i, I-c-ii, II, II-a, II-b,and II-c, as set forth below.

In some embodiments, one or more compounds of Formula I is providedand/or utilized in a solid form (e.g., a crystal form or an amorphousform).

In some embodiments, the present disclosure provides compositions thatcomprise and/or deliver a compound of Formula I (e.g., in a form asdescribed herein), a prodrug or active metabolite thereof.

In some embodiments, the present disclosure provides compositions thatcomprise and/or deliver a compound of Formula I. In some embodiments,such compositions are pharmaceutical compositions that include at leastone pharmaceutically acceptable carrier, diluent or excipient.

In some embodiments, provided SARM1 inhibitors reduce or inhibit bindingof NAD+ by SARM1. In some embodiments, provided SARM1 inhibitors bind toSARM1 within a pocket comprising one or more catalytic residues (e.g., acatalytic cleft of SARM1).

In some embodiments, provided compounds and/or compositions inhibitactivity of SARM1. Alternatively or additionally, in some embodiments,provided compounds alleviate one or more attributes ofneurodegeneration. In some embodiments, the present disclosure providesmethods of treating a neurodegenerative disease or disorder associatedwith axonal degeneration.

In some embodiments, one or more compounds and/or compositions asdescribed herein are useful, for example, in the practice of medicine.In some embodiments, one or more compounds and/or compositions asdescribed herein are useful, for example, to treat, prevent, orameliorate axonal degeneration (e.g., one or more features orcharacteristics thereof). In some embodiments, one or more compoundsand/or compositions as described herein are useful, for example, toinhibit axonal degeneration, including axonal degeneration that resultsfrom reduction or depletion of NAD+. In some embodiments, one or morecompounds and/or compositions as described herein are useful, forexample, to prevent the axon distal to an axonal injury fromdegenerating.

In some embodiments, one or more compounds and/or compositions asdescribed herein are useful, for example, to treat one or moreneurodegenerative diseases, disorders or conditions selected from thegroup consisting of neuropathies or axonopathies. In some embodiments,one or more compounds and/or compositions as described herein areuseful, for example, to treat a neuropathy or axonopathy associated withaxonal degeneration. In some embodiments, a neuropathy associated withaxonal degeneration is a hereditary or congenital neuropathy oraxonopathy. In some embodiments, a neuropathy associated with axonaldegeneration results from a de novo or somatic mutation. In someembodiments, a neuropathy associated with axonal degeneration isselected from a list contained herein. In some embodiments, a neuropathyor axonopathy is associated with axonal degeneration, including, but notlimited to, chemotherapy-induced peripheral neuropathy, multiplesclerosis, optic neuropathies such as glaucoma, Parkinson's disease,Alzheimer's disease, Herpes infection, diabetes, amyotrophic lateralsclerosis (ALS), a demyelinating disease, ischemia or stroke, chemicalinjury, thermal injury, and AIDS.

In some embodiments, subjects to which a compound or composition asdescribed herein is administered may be or comprise subjects sufferingfrom or susceptible to a neurodegenerative disease, disorder orcondition. In some embodiments, a neurodegenerative disease, disorder orcondition may be or comprise a traumatic neuronal injury. In someembodiments, a traumatic neuronal injury is blunt force trauma, aclosed-head injury, an open head injury, exposure to a concussive and/orexplosive force, a penetrating injury in or to the brain cavity orinnervated region of the body. In some embodiments, a traumatic neuronalinjury is a force which causes axons to deform, stretch, crush or sheer.

In some embodiments, provided methods comprise administering a compounddescribed herein to a patient in need thereof. In some such embodiments,the patient is at risk of developing a condition characterized by axonaldegeneration. In some embodiments, the patient has a conditioncharacterized by axonal degeneration. In some embodiments, the patienthas been diagnosed with a condition characterized by axonaldegeneration.

In some embodiments, provided methods comprise administering acomposition as described herein to a patient population in need thereof.In some embodiments, the population is drawn from individuals who engagein activities where the potential for traumatic neuronal injury is high.In some embodiments, the population is drawn from athletes who engage incontact sports or other high-risk activities

In some embodiments, the patient is at risk of developing aneurodegenerative disorder. In some embodiments the patient is elderly.In some embodiments, the patient is known to have a genetic risk factorfor neurodegeneration.

In certain embodiments, the present disclosure provides compounds thatare useful, for example, as analytical tools, as probes in biologicalassays, or as therapeutic agents in accordance with the presentdisclosure. Compounds provided by this disclosure are also useful forthe study of SARM1 function in biological and pathological phenomena andthe comparative evaluation of new SARM1 activity inhibitors in vitro orin vivo.

In some embodiments, one or more compounds and/or compositions asdescribed herein are useful, for example, as a method for inhibiting thedegradation of neurons derived from a subject. In some embodiments, oneor more compounds and/or compositions as described herein are useful forinhibiting the degeneration of a neuron, or a portion thereof, culturedin vitro. In some embodiments, one or more compounds and/or compositionsas described herein are useful as stabilizing agents to promote in vitroneuronal survival.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 illustrates the structure of the SARM1 protein.

DEFINITIONS

Aliphatic: The term “aliphatic” refers to a straight-chain (i.e.,unbranched) or branched, substituted or unsubstituted hydrocarbon chainthat is completely saturated or that contains one or more units ofunsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon thatis completely saturated or that contains one or more units ofunsaturation, but which is not aromatic (also referred to herein as“carbocycle” or “cycloaliphatic”), that has a single point of attachmentto the rest of the molecule. Unless otherwise specified, aliphaticgroups contain 1-6 aliphatic carbon atoms. In some embodiments,aliphatic groups contain 1-5 aliphatic carbon atoms. In otherembodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. Instill other embodiments, aliphatic groups contain 1-3 aliphatic carbonatoms, and in yet other embodiments, aliphatic groups contain 1-2aliphatic carbon atoms. In some embodiments, “cycloaliphatic” (or“carbocycle”) refers to a monocyclic C₃-C₈ hydrocarbon or a bicyclicC₇-C₁₀ hydrocarbon that is completely saturated or that contains one ormore units of unsaturation, but which is not aromatic, that has a singlepoint of attachment to the rest of the molecule. Suitable aliphaticgroups include, but are not limited to, linear or branched, substitutedor unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof.

Alkyl: The term “alkyl”, used alone or as part of a larger moiety,refers to a saturated, optionally substituted straight or branched chainor cyclic hydrocarbon group having 1-12, 1-10, 1-8, 1-6, 1-4, 1-3, or1-2 carbon atoms. The term “cycloalkyl” refers to an optionallysubstituted saturated ring system of about 3 to about 10 ring carbonatoms. Exemplary monocyclic cycloalkyl rings include cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

Alkylene: The term “alkylene” refers to a bivalent alkyl group. In someembodiments, “alkylene” is a bivalent straight or branched alkyl group.In some embodiments, an “alkylene chain” is a polymethylene group, i.e.,—(CH₂)_(n)—, wherein n is a positive integer, e.g., from 1 to 6, from 1to 4, from 1 to 3, from 1 to 2, or from 2 to 3. An optionallysubstituted alkylene chain is a polymethylene group in which one or moremethylene hydrogen atoms is optionally replaced with a substituent.Suitable substituents include those described below for a substitutedaliphatic group and also include those described in the specificationherein. It will be appreciated that two substituents of the alkylenegroup may be taken together to form a ring system. In certainembodiments, two substituents can be taken together to form a 3- to7-membered ring. The substituents can be on the same or different atoms.

Alkenyl: The term “alkenyl”, used alone or as part of a larger moiety,refers to an optionally substituted straight or branched chain or cyclichydrocarbon group having at least one double bond and having 2-12, 2-10,2-8, 2-6, 2-4, or 2-3 carbon atoms. The term “cycloalkenyl” refers to anoptionally substituted non-aromatic monocyclic or multicyclic ringsystem containing at least one carbon-carbon double bond and havingabout 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkenylrings include cyclopentyl, cyclohexenyl, and cycloheptenyl.

Alkynyl: The term “alkynyl”, used alone or as part of a larger moiety,refers to an optionally substituted straight or branched chainhydrocarbon group having at least one triple bond and having 2-12, 2-10,2-8, 2-6, 2-4, or 2-3 carbon atoms.

Aryl: The term “aryl” refers to monocyclic and bicyclic ring systemshaving a total of five to fourteen ring members, wherein at least onering in the system is aromatic and wherein each ring in the systemcontains three to seven ring members. The term “aryl” may be usedinterchangeably with the term “aryl ring”. In certain embodiments of thepresent invention, “aryl” refers to an aromatic ring system whichincludes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl andthe like, which may bear one or more substituents. Also included withinthe scope of the term “aryl”, as it is used herein, is a group in whichan aromatic ring is fused to one or more non-aromatic carbocyclic orheterocyclic rings, such as indanyl, phthalimidyl, naphthimidyl,phenanthridinyl, tetrahydronaphthyl, imidazolidinyl, imidazolidin-2-one,and the like.

Binding: It will be understood that the term “binding”, as used herein,typically refers to a non-covalent association between or among two ormore entities. “Direct” binding involves physical contact betweenentities or moieties; indirect binding involves physical interaction byway of physical contact with one or more intermediate entities. Bindingbetween two or more entities can typically be assessed in any of avariety of contexts—including where interacting entities or moieties arestudied in isolation or in the context of more complex systems (e.g.,while covalently or otherwise associated with a carrier entity and/or ina biological system or cell).

Biological Sample: As used herein, the term “biological sample”typically refers to a sample obtained or derived from a biologicalsource (e.g., a tissue or organism or cell culture) of interest, asdescribed herein. In some embodiments, a source of interest comprises anorganism, such as an animal or human. In some embodiments, a biologicalsample is or comprises biological tissue or fluid. In some embodiments,a biological sample may be or comprise bone marrow; blood; blood cells;ascites; tissue or fine needle biopsy samples; cell-containing bodyfluids; free floating nucleic acids; sputum; saliva; urine;cerebrospinal fluid, peritoneal fluid; pleural fluid; feces; lymph;gynecological fluids; skin swabs; vaginal swabs; oral swabs; nasalswabs; washings or lavages such as a ductal lavages or broncheoalveolarlavages; aspirates; scrapings; bone marrow specimens; tissue biopsyspecimens; surgical specimens; feces, other body fluids, secretions,and/or excretions; and/or cells therefrom, etc. In some embodiments, abiological sample is or comprises cells obtained from an individual. Insome embodiments, obtained cells are or include cells from an individualfrom whom the sample is obtained. In some embodiments, a sample is a“primary sample” obtained directly from a source of interest by anyappropriate means. For example, in some embodiments, a primarybiological sample is obtained by methods selected from the groupconsisting of biopsy (e.g., fine needle aspiration or tissue biopsy),surgery, collection of body fluid (e.g., blood, lymph, feces etc.), etc.In some embodiments, as will be clear from context, the term “sample”refers to a preparation that is obtained by processing (e.g., byremoving one or more components of and/or by adding one or more agentsto) a primary sample. For example, filtering using a semi-permeablemembrane. Such a “processed sample” may comprise, for example, nucleicacids or proteins extracted from a sample or obtained by subjecting aprimary sample to techniques such as amplification or reversetranscription of mRNA, isolation and/or purification of certaincomponents, etc.

Biomarker: The term “biomarker” is used herein to refer to a to anentity, event, or characteristic whose presence, level, degree, type,and/or form, correlates with a particular biological event or state ofinterest, so that it is considered to be a “marker” of that event orstate. To give but a few examples, in some embodiments, a biomarker maybe or comprise a marker for a particular disease state, or forlikelihood that a particular disease, disorder or condition may develop,occur, or reoccur. In some embodiments, a biomarker may be or comprise amarker for a particular disease or therapeutic outcome, or likelihoodthereof. Thus, in some embodiments, a biomarker is predictive, in someembodiments, a biomarker is prognostic, in some embodiments, a biomarkeris diagnostic, of the relevant biological event or state of interest. Abiomarker may be or comprise an entity of any chemical class, and may beor comprise a combination of entities. For example, in some embodiments,a biomarker may be or comprise a nucleic acid, a polypeptide, a lipid, acarbohydrate, a small molecule, an inorganic agent (e.g., a metal orion), or a combination thereof. In some embodiments, a biomarker is acell surface marker. In some embodiments, a biomarker is intracellular.In some embodiments, a biomarker is detected outside of cells (e.g., issecreted or is otherwise generated or present outside of cells, e.g., ina body fluid such as blood, urine, tears, saliva, cerebrospinal fluid,etc. In some embodiments, a biomarker may be or comprise a genetic orepigenetic signature. In some embodiments, a biomarker may be orcomprise a gene expression signature.

In some embodiments, a biomarker may be or comprise a marker forneurodegeneration, or for likelihood that a neurodegenerative disease,disorder or condition may develop, occur, or reoccur. In someembodiments, a biomarker may be or comprise a marker ofneurodegeneration a therapeutic outcome, or likelihood thereof. Thus, insome embodiments, a biomarker is predictive, in some embodiments, abiomarker is prognostic, in some embodiments, a biomarker is diagnostic,of a neurodegenerative disease, disorder or condition. In someembodiments changes in biomarker levels can be detected via cerebralspinal fluid (CSF), plasma and/or serum.

In some embodiments, neurodegeneration may be assessed, for example, bydetecting an increase and/or decrease in the concentration ofneurofilament protein light (NF-L) and/or neurofilament protein heavy(NF-H) contained the cerebral spinal fluid of a subject. In someembodiments, the incidence and/or progression of neurodegeneration canbe assessed via positron emission tomography (PET) with a synapticvesicle glycoprotein 2a (SV2A) ligand. In some embodiments, a detectablechange in constitutive NAD and/or cADPR levels in neurons can be used toassess neurodegeneration.

In some embodiments, a detectable change in one or moreneurodegeneration associated proteins in a subject, relative to ahealthy reference population can be used as a biomarker ofneurodegeneration. Such proteins include, but are not limited to,albumin, amyloid-β (Aβ)38, Aβ40, Aβ42, glial fibrillary acid protein(GFAP), heart-type fatty acid binding protein (hFABP), monocytechemoattractin protein (MCP)-1, neurogranin, neuron specific enolayse(NSE), soluble amyloid precursor protein (sAPP)α, sAPPβ, solubletriggering receptor expressed on myeloid cells (sTREM) 2, phospho-tau,and/or total-tau. In some embodiments, an increase in cytokines and/orchemokines, including, but not limited to, Ccl2, Ccl7, Ccl12, Csf1,and/or Il6, can be used as a biomarker of neurodegeneration.

Carrier: As used herein, the term “carrier” refers to a diluent,adjuvant, excipient, or vehicle with which a composition isadministered. In some exemplary embodiments, carriers can includesterile liquids, such as, for example, water and oils, including oils ofpetroleum, animal, vegetable or synthetic origin, such as, for example,peanut oil, soybean oil, mineral oil, sesame oil and the like. In someembodiments, carriers are or include one or more solid components.

Combination therapy: As used herein, the term “combination therapy”refers to those situations in which a subject is simultaneously exposedto two or more therapeutic regimens (e.g., two or more therapeuticagents). In some embodiments, the two or more regimens may beadministered simultaneously; in some embodiments, such regimens may beadministered sequentially (e.g., all “doses” of a first regimen areadministered prior to administration of any doses of a second regimen);in some embodiments, such agents are administered in overlapping dosingregimens. In some embodiments, “administration” of combination therapymay involve administration of one or more agent(s) or modality(ies) to asubject receiving the other agent(s) or modality(ies) in thecombination. For clarity, combination therapy does not require thatindividual agents be administered together in a single composition (oreven necessarily at the same time), although in some embodiments, two ormore agents, or active moieties thereof, may be administered together ina combination composition, or even in a combination compound (e.g., aspart of a single chemical complex or covalent entity).

Composition: Those skilled in the art will appreciate that the term“composition” may be used to refer to a discrete physical entity thatcomprises one or more specified components. In general, unless otherwisespecified, a composition may be of any form—e.g., gas, gel, liquid,solid, etc.

Domain: The term “domain” as used herein refers to a section or portionof an entity. In some embodiments, a “domain” is associated with aparticular structural and/or functional feature of the entity so that,when the domain is physically separated from the rest of its parententity, it substantially or entirely retains the particular structuraland/or functional feature. Alternatively or additionally, a domain maybe or include a portion of an entity that, when separated from that(parent) entity and linked with a different (recipient) entity,substantially retains and/or imparts on the recipient entity one or morestructural and/or functional features that characterized it in theparent entity. In some embodiments, a domain is a section or portion ofa molecule (e.g., a small molecule, carbohydrate, lipid, nucleic acid,or polypeptide). In some embodiments, a domain is a section of apolypeptide; in some such embodiments, a domain is characterized by aparticular structural element (e.g., a particular amino acid sequence orsequence motif, α-helix character, β-sheet character, coiled-coilcharacter, random coil character, etc.), and/or by a particularfunctional feature (e.g., binding activity, enzymatic activity, foldingactivity, signaling activity, etc.).

Dosage form or unit dosage form: Those skilled in the art willappreciate that the term “dosage form” may be used to refer to aphysically discrete unit of an active agent (e.g., a therapeutic ordiagnostic agent) for administration to a subject. Typically, each suchunit contains a predetermined quantity of active agent. In someembodiments, such quantity is a unit dosage amount (or a whole fractionthereof) appropriate for administration in accordance with a dosingregimen that has been determined to correlate with a desired orbeneficial outcome when administered to a relevant population (i.e.,with a therapeutic dosing regimen). Those of ordinary skill in the artappreciate that the total amount of a therapeutic composition or agentadministered to a particular subject is determined by one or moreattending physicians and may involve administration of multiple dosageforms.

Dosing regimen or therapeutic regimen: Those skilled in the art willappreciate that the terms “dosing regimen” and “therapeutic regimen” maybe used to refer to a set of unit doses (typically more than one) thatare administered individually to a subject, typically separated byperiods of time. In some embodiments, a given therapeutic agent has arecommended dosing regimen, which may involve one or more doses. In someembodiments, a dosing regimen comprises a plurality of doses each ofwhich is separated in time from other doses. In some embodiments,individual doses are separated from one another by a time period of thesame length; in some embodiments, a dosing regimen comprises a pluralityof doses and at least two different time periods separating individualdoses. In some embodiments, all doses within a dosing regimen are of thesame unit dose amount. In some embodiments, different doses within adosing regimen are of different amounts. In some embodiments, a dosingregimen comprises a first dose in a first dose amount, followed by oneor more additional doses in a second dose amount different from thefirst dose amount. In some embodiments, a dosing regimen comprises afirst dose in a first dose amount, followed by one or more additionaldoses in a second dose amount same as the first dose amount. In someembodiments, a dosing regimen is correlated with a desired or beneficialoutcome when administered across a relevant population (i.e., is atherapeutic dosing regimen).

Excipient: as used herein, refers to a non-therapeutic agent that may beincluded in a pharmaceutical composition, for example, to provide orcontribute to a desired consistency or stabilizing effect. Suitablepharmaceutical excipients include, for example, starch, glucose,lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodiumstearate, glycerol monostearate, talc, sodium chloride, dried skim milk,glycerol, propylene, glycol, water, ethanol and the like.

Heteroaryl: The terms “heteroaryl” and “heteroar-”, used alone or aspart of a larger moiety, e.g., “heteroaralkyl”, or “heteroaralkoxy”,refer to groups having 5 to 10 ring atoms, preferably 5, 6, 9, or 10ring atoms; having 6, 10, or 14 π electrons shared in a cyclic array;and having, in addition to carbon atoms, from one to five heteroatoms.The term “heteroatom” refers to nitrogen, oxygen, or sulfur, andincludes any oxidized form of nitrogen or sulfur, and any quaternizedform of a basic nitrogen. Heteroaryl groups include, without limitation,thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl,tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl,thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl,purinyl, naphthyridinyl, and pteridinyl. The terms “heteroaryl” and“heteroar-”, as used herein, also include groups in which aheteroaromatic ring is fused to one or more aryl, cycloaliphatic, orheterocyclyl rings, where the radical or point of attachment is on theheteroaromatic ring. Nonlimiting examples include indolyl, isoindolyl,benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl,benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl,quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl,phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Aheteroaryl group may be mono- or bicyclic. The term “heteroaryl” may beused interchangeably with the terms “heteroaryl ring”, “heteroarylgroup”, or “heteroaromatic”, any of which terms include rings that areoptionally substituted. The term “heteroaralkyl” refers to an alkylgroup substituted by a heteroaryl, wherein the alkyl and heteroarylportions independently are optionally substituted.

Heterocycle: As used herein, the terms “heterocycle”, “heterocyclyl”,“heterocyclic radical”, and “heterocyclic ring” are used interchangeablyand refer to a stable 3- to 8-membered monocyclic or 7-10-memberedbicyclic heterocyclic moiety that is either saturated or partiallyunsaturated, and having, in addition to carbon atoms, one or more, suchas one to four, heteroatoms, as defined above. When used in reference toa ring atom of a heterocycle, the term “nitrogen” includes a substitutednitrogen. As an example, in a saturated or partially unsaturated ringhaving 0-3 heteroatoms selected from oxygen, sulfur and nitrogen, thenitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as inpyrrolidinyl), or NR⁺ (as in N-substituted pyrrolidinyl). A heterocyclicring can be attached to its pendant group at any heteroatom or carbonatom that results in a stable structure and any of the ring atoms can beoptionally substituted. Examples of such saturated or partiallyunsaturated heterocyclic radicals include, without limitation,tetrahydrofuranyl, tetrahydrothienyl, piperidinyl, decahydroquinolinyl,oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl,thiazepinyl, morpholinyl, and thiamorpholinyl. A heterocyclyl group maybe mono-, bi-, tri-, or polycyclic, preferably mono-, bi-, or tricyclic,more preferably mono- or bicyclic. The term “heterocyclylalkyl” refersto an alkyl group substituted by a heterocyclyl, wherein the alkyl andheterocyclyl portions independently are optionally substituted.Additionally, a heterocyclic ring also includes groups in which theheterocyclic ring is fused to one or more aryl rings (e.g.,2,3-dihydrobenzofuran, 2,3-dihydrobenzo[b][1,4]dioxine, etc.).

Inhibitory agent: As used herein, the term “inhibitory agent” refers toan entity, condition, or event whose presence, level, or degreecorrelates with decreased level or activity of a target. In someembodiments, an inhibitory agent may act directly (in which case itexerts its influence directly upon its target, for example, by bindingto the target); in some embodiments, an inhibitory agent may actindirectly (in which case it exerts its influence by interacting withand/or otherwise altering a regulator of the target, so that leveland/or activity of the target is reduced). In some embodiments, aninhibitory agent is one whose presence or level correlates with a targetlevel or activity that is reduced relative to a particular referencelevel or activity (e.g., that observed under appropriate referenceconditions, such as presence of a known inhibitory agent, or absence ofthe inhibitory agent in question, etc.).

Neurodegeneration: As used herein, the term “neurodegeneration” refersto a reduction in one or more features, structures, or characteristicsof a neuron or neuronal tissue. In some embodiments, neurodegenerationis observed as a pathological reduction in an organism. Those skilled inthe art will appreciate that neurodegeneration is associated withcertain diseases, disorders and conditions, including those that affecthumans. In some embodiments, neurodegeneration may be transient (e.g.,as sometimes occurs in association with certain infections and/orchemical or mechanical disruptions); in some embodiments,neurodegeneration may be chronic and/or progressive (e.g., as is oftenassociated with certain diseases, disorders or conditions such as, butnot limited to, Parkinson's disease, amyotrophic lateral sclerosis,multiple sclerosis, Huntington disease, or Alzheimer's disease). In someembodiments, neurodegeneration may be assessed, for example, bydetecting in a subject an increase in a biomarker associated withneurodegeneration. In some embodiments, neurodegeneration may beassessed, for example, by detecting in a subject a decrease in abiomarker associated with neurodegeneration. Alternatively oradditionally, in some embodiments, neurodegeneration may be assessed bymagnetic resonance imaging (MRI), biomarkers containing cerebral spinalfluid, or other biomarkers observed in patients. In some embodiments,neurodegeneration is defined as a score of below 24 on the mini-mentalstate examination. In some embodiments, neurodegeneration refers to lossof synapses. In some embodiments, neurodegeneration refers to areduction in neural tissue relating to a traumatic injury (e.g. exposureto an external force which disrupts the integrity of the neural tissue).In some embodiments, neurodegeneration refers to a reduction inperipheral neural tissue. In some embodiments, neurodegeneration refersto a reduction in central nervous tissue.

Oral: The phrases “oral administration” and “administered orally” asused herein have their art-understood meaning referring toadministration by mouth of a compound or composition.

Parenteral: The phrases “parenteral administration” and “administeredparenterally” as used herein have their art-understood meaning referringto modes of administration other than enteral and topicaladministration, usually by injection, and include, without limitation,intravenous, intramuscular, intra-arterial, intrathecal, intracapsular,intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal,subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid,intraspinal, and intrasternal injection and infusion.

Partially Unsaturated: As used herein, the term “partially unsaturated”refers to a ring moiety that includes at least one double or triple bondbetween ring atoms. The term “partially unsaturated” is intended toencompass rings having multiple sites of unsaturation, but is notintended to include aromatic (e.g., aryl or heteroaryl) moieties, asherein defined.

Patient: As used herein, the term “patient” refers to any organism towhich a provided composition is or may be administered, e.g., forexperimental, diagnostic, prophylactic, cosmetic, and/or therapeuticpurposes. Typical patients include animals (e.g., mammals such as mice,rats, rabbits, non-human primates, and/or humans). In some embodiments,a patient is a human. In some embodiments, a patient is suffering fromor susceptible to one or more disorders or conditions. In someembodiments, a patient displays one or more symptoms of a disorder orcondition. In some embodiments, a patient has been diagnosed with one ormore disorders or conditions. In some embodiments, the patient isreceiving or has received certain therapy to diagnose and/or to treat adisease, disorder, or condition.

Pharmaceutical composition: As used herein, the term “pharmaceuticalcomposition” refers to an active agent, formulated together with one ormore pharmaceutically acceptable carriers. In some embodiments, theactive agent is present in unit dose amount appropriate foradministration in a therapeutic or dosing regimen that shows astatistically significant probability of achieving a predeterminedtherapeutic effect when administered to a relevant population. In someembodiments, pharmaceutical compositions may be specially formulated foradministration in solid or liquid form, including those adapted for thefollowing: oral administration, for example, drenches (aqueous ornon-aqueous solutions or suspensions), tablets, e.g., those targeted forbuccal, sublingual, and systemic absorption, boluses, powders, granules,pastes for application to the tongue; parenteral administration, forexample, by subcutaneous, intramuscular, intravenous or epiduralinjection as, for example, a sterile solution or suspension, orsustained-release formulation; topical application, for example, as acream, ointment, or a controlled-release patch or spray applied to theskin, lungs, or oral cavity; intravaginally or intrarectally, forexample, as a pessary, cream, or foam; sublingually; ocularly;transdermally; or nasally, pulmonary, and to other mucosal surfaces.

Pharmaceutically acceptable: As used herein, the phrase“pharmaceutically acceptable” refers to those compounds, materials,compositions, and/or dosage forms which are, within the scope of soundmedical judgment, suitable for use in contact with the tissues of humanbeings and animals without excessive toxicity, irritation, allergicresponse, or other problem or complication, commensurate with areasonable benefit/risk ratio.

Pharmaceutically acceptable carrier: As used herein, the term“pharmaceutically acceptable carrier” means apharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, or solvent encapsulatingmaterial, involved in carrying or transporting the subject compound fromone organ, or portion of the body, to another organ, or portion of thebody. Each carrier must be “acceptable” in the sense of being compatiblewith the other ingredients of the formulation and not injurious to thepatient. Some examples of materials which can serve aspharmaceutically-acceptable carriers include: sugars, such as lactose,glucose and sucrose; starches, such as corn starch and potato starch;cellulose, and its derivatives, such as sodium carboxymethyl cellulose,ethyl cellulose and cellulose acetate; powdered tragacanth; malt;gelatin; talc; excipients, such as cocoa butter and suppository waxes;oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil,olive oil, corn oil and soybean oil; glycols, such as propylene glycol;polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol;esters, such as ethyl oleate and ethyl laurate; agar; buffering agents,such as magnesium hydroxide and aluminum hydroxide; alginic acid;pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol;pH buffered solutions; polyesters, polycarbonates and/or polyanhydrides;and other nontoxic compatible substances employed in pharmaceuticalformulations.

Pharmaceutically acceptable salt: The term “pharmaceutically acceptablesalt”, as used herein, refers to salts of such compounds that areappropriate for use in pharmaceutical contexts, i.e., salts which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and lower animals without undue toxicity,irritation, allergic response and the like, and are commensurate with areasonable benefit/risk ratio. Pharmaceutically acceptable salts arewell known in the art. For example, S. M. Berge, et al. describespharmaceutically acceptable salts in detail in J. PharmaceuticalSciences, 66: 1-19 (1977). In some embodiments, pharmaceuticallyacceptable salts include, but are not limited to, nontoxic acid additionsalts, which are salts of an amino group formed with inorganic acidssuch as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuricacid and perchloric acid or with organic acids such as acetic acid,maleic acid, tartaric acid, citric acid, succinic acid or malonic acidor by using other methods used in the art such as ion exchange. In someembodiments, pharmaceutically acceptable salts include, but are notlimited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate,benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate,citrate, cyclopentanepropionate, digluconate, dodecylsulfate,ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate,gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Representative alkali or alkaline earth metal salts includesodium, lithium, potassium, calcium, magnesium, and the like. In someembodiments, pharmaceutically acceptable salts include, whenappropriate, nontoxic ammonium, quaternary ammonium, and amine cationsformed using counterions such as halide, hydroxide, carboxylate,sulfate, phosphate, nitrate, alkyl having from 1 to 6 carbon atoms,sulfonate and aryl sulfonate.

Prevent or prevention: As used herein, the terms “prevent” or“prevention”, when used in connection with the occurrence of a disease,disorder, and/or condition, refer to reducing the risk of developing thedisease, disorder and/or condition and/or to delaying onset of one ormore characteristics or symptoms of the disease, disorder or condition.Prevention may be considered complete when onset of a disease, disorderor condition has been delayed for a predefined period of time.

Specific: The term “specific”, when used herein with reference to anagent having an activity, is understood by those skilled in the art tomean that the agent discriminates between potential target entities orstates. For example, in some embodiments, an agent is said to bind“specifically” to its target if it binds preferentially with that targetin the presence of one or more competing alternative targets. In manyembodiments, specific interaction is dependent upon the presence of aparticular structural feature of the target entity (e.g., an epitope, acleft, a binding site). It is to be understood that specificity need notbe absolute. In some embodiments, specificity may be evaluated relativeto that of the binding agent for one or more other potential targetentities (e.g., competitors). In some embodiments, specificity isevaluated relative to that of a reference specific binding agent. Insome embodiments, specificity is evaluated relative to that of areference non-specific binding agent. In some embodiments, the agent orentity does not detectably bind to the competing alternative targetunder conditions of binding to its target entity. In some embodiments, abinding agent binds with higher on-rate, lower off-rate, increasedaffinity, decreased dissociation, and/or increased stability to itstarget entity as compared with the competing alternative target(s).

Subject: As used herein, the term “subject” refers to an organism,typically a mammal (e.g., a human, in some embodiments includingprenatal human forms). In some embodiments, a subject is suffering froma relevant disease, disorder or condition. In some embodiments, asubject is susceptible to a disease, disorder, or condition. In someembodiments, a subject displays one or more symptoms or characteristicsof a disease, disorder or condition. In some embodiments, a subject doesnot display any symptom or characteristic of a disease, disorder, orcondition. In some embodiments, a subject is someone with one or morefeatures characteristic of susceptibility to or risk of a disease,disorder, or condition. In some embodiments, a subject is a patient. Insome embodiments, a subject is an individual to whom diagnosis and/ortherapy is and/or has been administered.

Substituted or optionally substituted: As described herein, compounds ofthe invention may contain “optionally substituted” moieties. In general,the term “substituted,” whether preceded by the term “optionally” ornot, means that one or more hydrogens of the designated moiety arereplaced with a suitable substituent. “Substituted” applies to one ormore hydrogens that are either explicit or implicit from the structure(e.g.,

refers to at least

refers to at least

Unless otherwise indicated, an “optionally substituted” group may have asuitable substituent at each substitutable position of the group, andwhen more than one position in any given structure may be substitutedwith more than one substituent selected from a specified group, thesubstituent may be either the same or different at every position.Combinations of substituents envisioned by this invention are preferablythose that result in the formation of stable or chemically feasiblecompounds. The term “stable,” as used herein, refers to compounds thatare not substantially altered when subjected to conditions to allow fortheir production, detection, and, in certain embodiments, theirrecovery, purification, and use for one or more of the purposesdisclosed herein.

Suitable monovalent substituents on a substitutable carbon atom of an“optionally substituted” group are independently halogen;—(CH₂)₀₋₄R^(∘); —(CH₂)₀₋₄OR^(∘); —O(CH₂)₀₋₄R^(∘), —O—(CH₂)₀₋₄C(O)OR^(∘);—(CH₂)₀₋₄CH(OR^(∘))₂; —(CH₂)₀₋₄SR^(∘); —(CH₂)₀₋₄Ph, which may besubstituted with R^(∘); —(CH₂)₀₋₄O(CH₂)₀₋₁Ph which may be substitutedwith R^(∘); —CH═CHPh, which may be substituted with R^(∘);—(CH₂)₀₋₄O(CH₂)₀₋₁-pyridyl which may be substituted with R^(∘); —NO₂;—CN; —N₃; —(CH₂)₀₋₄N(R^(∘))₂; —(CH₂)₀₋₄N(R^(∘))C(O)R^(∘);—N(R^(∘))C(S)R^(∘); —(CH₂)₀₋₄N(R^(∘))C(O)NR^(∘) ₂; —N(R^(∘))C(S)NR^(∘)₂; —(CH₂)₀₋₄N(R^(∘))C(O)OR^(∘); —N(R^(∘))N(R^(∘))C(O)R^(∘);—N(R^(∘))N(R^(∘))C(O)NR^(∘) ₂; —N(R^(∘))N(R^(∘))C(O)OR^(∘);—(CH₂)₀₋₄C(O)R^(∘); —C(S)R^(∘); —(CH₂)₀₋₄C(O)OR^(∘);—(CH₂)₀₋₄C(O)SR^(∘); —(CH₂)₀₋₄C(O)OSiR^(∘) ₃; —(CH₂)₀₋₄OC(O)R^(∘);—OC(O)(CH₂)₀₋₄SR^(∘); —(CH₂)₀₋₄SC(O)R^(∘); —(CH₂)₀₋₄C(O)NR^(∘) ₂;—C(S)NR^(∘) ₂; —C(S)SR^(∘); —SC(S)SR^(∘), —(CH₂)₀₋₄OC(O)NR^(∘) ₂;—C(O)N(OR^(∘))R^(∘); —C(O)C(O)R^(∘); —C(O)CH₂C(O)R^(∘);—C(NOR^(∘))R^(∘); —(CH₂)₀₋₄SSR^(∘); —(CH₂)₀₋₄S(O)₂R^(∘);—(CH₂)₀₋₄S(O)(NH)R^(∘); —(CH₂)₀₋₄S(O)₂OR^(∘); —(CH₂)₀₋₄OS(O)₂R^(∘);—S(O)₂NR^(∘) ₂; —(CH₂)₀₋₄S(O)R^(∘); —N(R^(∘))S(O)₂NR^(∘) ₂;—N(R^(∘))S(O)₂R^(∘); —N(OR^(∘))R^(∘); —C(NH)NR^(∘) ₂; —P(O)₂R^(∘);—P(O)R^(∘) ₂; —OP(O)R^(∘) ₂; —OP(O)(OR^(∘))₂; SiR^(∘) ₃; —(C₁₋₄ straightor branched alkylene)O—N(R^(∘))₂; or —(C₁₋₄ straight or branchedalkylene)C(O)O—N(R^(∘))₂, wherein each R^(∘) may be substituted asdefined below and is independently hydrogen, C₁₋₆ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, —CH₂-(5- to 6-membered heteroaryl ring), a 5- to6-membered saturated, partially unsaturated, or aryl ring having 0-4heteroatoms independently selected from nitrogen, oxygen, or sulfur, oran 8- to 10-membered bicyclic aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences ofR^(∘), taken together with their intervening atom(s), form a 3- to12-membered saturated, partially unsaturated, or aryl mono- or bicyclicring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur, which may be substituted as defined below.

Suitable monovalent substituents on R^(∘) (or the ring formed by takingtwo independent occurrences of R^(∘) together with their interveningatoms), are independently halogen, —(CH₂)₀₋₂R^(●), -(haloR^(●)),—(CH₂)₀₋₂OH, —(CH₂)₀₋₂OR^(●), —(CH₂)₀₋₂CH(OR^(●))₂; —O(haloR^(●)), —CN,—N₃, —(CH₂)₀₋₂C(O)R^(●)—(CH₂)₀₋₂C(O)OH, —(CH₂)₀₋₂C(O)OR^(●),—(CH₂)₀₋₂SR^(●), —(CH₂)₀₋₂SH, —(CH₂)₀₋₂NH₂, —(CH₂)₀₋₂NHR^(●),—(CH₂)₀₋₂NR^(●) ₂, —NO₂, —SiR^(●) ₃, —OSiR^(●) ₃, —C(O)SR^(●), —(C₁₋₄straight or branched alkylene)C(O)OR^(●), or —SSR^(●) wherein each R^(●)is unsubstituted or where preceded by “halo” is substituted only withone or more halogens, and is independently selected from C₁₋₄ aliphatic,—CH₂Ph, —O(CH₂)₀₋₁Ph, or a 3- to 6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. Suitable divalent substituents on asaturated carbon atom of R^(∘) include ═O and ═S.

Suitable divalent substituents on a saturated carbon atom of an“optionally substituted” group include the following: ═O (“oxo”), ═S,═NNR*2, ═NNHC(O)R*, ═NNHC(O)OR*, ═NNHS(O)₂R*, ═NR*, ═NOR*,—O(C(R*₂))₂₋₃O—, or —S(C(R*₂))₂₋₃S—, wherein each independent occurrenceof R* is selected from hydrogen, C₁₋₆ aliphatic which may be substitutedas defined below, or an unsubstituted 5- to 6-membered saturated,partially unsaturated, or aryl ring having 0-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur. Suitable divalentsubstituents that are bound to vicinal substitutable carbons of an“optionally substituted” group include: —O(CR*₂)₂₋₃O—, wherein eachindependent occurrence of R* is selected from hydrogen, C₁₋₆ aliphaticwhich may be substituted as defined below, or an unsubstituted5-6-membered saturated, partially unsaturated, or aryl ring having 0-4heteroatoms independently selected from nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R* include halogen,—R^(●), -(haloR^(●)), —OH, —OR^(●), —O(haloR^(●)), —CN, —C(O)OH,—C(O)OR^(●), —NH₂, —NHR^(●), —NR^(●) ₂, or —NO₂, wherein each R^(●) isunsubstituted or where preceded by “halo” is substituted only with oneor more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5- to 6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

Suitable substituents on a substitutable nitrogen of an “optionallysubstituted” group include —R^(†), —NR^(†) ₂, —C(O)R^(†), —C(O)OR^(†),—C(O)C(O)R^(†), —C(O)CH₂C(O)R^(†), —S(O)₂R^(†), —S(O)₂NR^(†) ₂,—C(S)NR^(†) ₂, —C(NH)NR^(†) ₂, or —N(R^(†))S(O)₂R^(†); wherein each R isindependently hydrogen, C₁₋₆ aliphatic which may be substituted asdefined below, unsubstituted —OPh, or an unsubstituted 5- to 6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences of R,taken together with their intervening atom(s) form an unsubstituted 3-to 12-membered saturated, partially unsaturated, or aryl mono- orbicyclic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R are independentlyhalogen, —R^(●), -(haloR^(●)), —OH, —OR^(●), —O(haloR^(●)), —CN,—C(O)OH, —C(O)OR^(●), —NH₂, —NHR^(●), —NR^(●) ₂, or —NO₂, wherein eachR^(●) is unsubstituted or where preceded by “halo” is substituted onlywith one or more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5- to 6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

Therapeutic agent: As used herein, the phrase “therapeutic agent” ingeneral refers to any agent that elicits a desired pharmacologicaleffect when administered to an organism. In some embodiments, an agentis considered to be a therapeutic agent if it demonstrates astatistically significant effect across an appropriate population. Insome embodiments, the appropriate population may be a population ofmodel organisms. In some embodiments, an appropriate population may bedefined by various criteria, such as a certain age group, gender,genetic background, preexisting clinical conditions, etc. In someembodiments, a therapeutic agent is a substance that can be used toalleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduceseverity of, and/or reduce incidence of one or more symptoms or featuresof a disease, disorder, and/or condition. In some embodiments, a“therapeutic agent” is an agent that has been or is required to beapproved by a government agency before it can be marketed foradministration to humans. In some embodiments, a “therapeutic agent” isan agent for which a medical prescription is required for administrationto humans.

Treat: As used herein, the terms “treat,” “treatment,” or “treating”refer to any method used to partially or completely alleviate,ameliorate, relieve, inhibit, prevent, delay onset of, reduce severityof, and/or reduce incidence of one or more symptoms or features of adisease, disorder, and/or condition. Treatment may be administered to asubject who does not exhibit signs of a disease, disorder, and/orcondition. In some embodiments, treatment may be administered to asubject who exhibits only early signs of the disease, disorder, and/orcondition, for example, for the purpose of decreasing the risk ofdeveloping pathology associated with the disease, disorder, and/orcondition.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS Programmed AxonalDegeneration and SARM1

Axonal degeneration is a major pathological feature of neurologicaldiseases such as, but not limited to, Alzheimer's disease, Parkinson'sdisease, ALS, multiple sclerosis, diabetic peripheral neuropathy,chemotherapy-induced peripheral neuropathy, inherited neuropathy,traumatic brain injury, and/or glaucoma. Damaged or unhealthy axons areeliminated via an intrinsic self-destruction program that is distinctfrom traditional cellular death pathways like apoptosis known asWallerian degeneration. (Gerdts, J., et al., Neuron, 2016, 89, 449-460;Whitmore, A. V. et al., Cell Death Differ., 2003, 10, 260-261). InWallerian degeneration, a peripheral nerve undergoes selective breakdownof the axon segment distal to an injury, whereas the proximal axonsegment and cell body remain intact. Without wishing to be bound bytheory, Wallerian degeneration is thought to be characterized, first, bya depletion of nicotinamide mononucleotide adenyltransferase (NMNAT),followed by nicotinamide adenine dinucleotide (NAD+) loss, adenosinetri-phosphate (ATP) loss, neurofilament proteolysis, and finally axonaldegradation approximately 8 to 24 hours following injury. (Gerdts, J.,et al., Neuron, 2016, 89, 449-460).

NAD+ is a ubiquitous metabolite with critical roles in energy metabolismand cell signaling (Belenkey et al., Trends Biochem., 2007, 32, 12-19;Chiarugi et al., Nat. Rev. Cancer, 2012, 12, 741-752). The homeostaticregulation of NAD+ levels is also responsible for maintaining axonalstability and integrity. (Babetto et al., Cell Rep., 2010, 3, 1422-1429;Sasaki et al., J. Neurosci., 2009).

In a genome-wide RNAi screen in primary mouse neurons, Sterile Alpha andTIR motif-containing 1 (SARM1) was identified, in which knockdown ofSARM1 led to long-lasting protection of sensory neurons againstinjury-induced axon degeneration (Gerdts et al., J Neurosci, 2013, 33,13569-13580). SARM1 belongs to the family of cytosolic adaptor proteins,but is unique among its members because it is the most evolutionaryancient adaptor, paradoxically inhibits TLR signaling, and has beenidentified as the central executioner of the injury-induced axon deathpathway (O'Neill, L. A. & Bowie, A. G., Nat. Rev. Immunol., 2007, 7,353-364; Osterloh, J. M., et al., Science, 2012, 337, 481-484; Gerdts,J., et al., J. Neurosci. 33, 2013, 13569-13580). Activation of SARM1 viaaxonal injury or forced dimerization of SARM1-TIR domains promotes rapidand catastrophic depletion of Nicotinamide Adenine Dinucleotide (NAD+),followed soon after by axonal degradation. (Gerdts, J., et al., Science,2015, 348, 453-457). SARM1 is required for this injury-induced NAD+depletion both in vitro and in vivo and SARM1 activation triggers axondegeneration locally via NAD(+) destruction (Gerdts et al., et al.,Science, 2015 348, 452-457; Sasaki et al., J. Biol. Chem. 2015, 290,17228-17238; both of which are hereby incorporated by reference in theirentireties).

From genetic loss-of-function studies it is clear that SARM1 serves asthe central executioner of the axonal degeneration pathway following aninjury. Genetic knockout of SARM1 allows for preservation of axons forup to 14 days after nerve transection (Osterloh, J. M., et al., Science,2012, 337, 481-484; Gerdts, J., et al., J. Neurosci., 2013, 33,13569-13580) and also improves functional outcomes in mice aftertraumatic brain injury (Henninger, N. et al., Brain 139, 2016,1094-1105). In addition to the role of SARM1 in direct axonal injury,SARM1 is also required for axonal degeneration observed inchemotherapy-induced peripheral neuropathy. Loss of SARM1 blockschemotherapy-induced peripheral neuropathy, both inhibiting axonaldegeneration and heightened pain sensitivity that develops afterchemotherapeutic vincristine treatment (Geisler et al, Brain, 2016, 139,3092-3108).

SARM1 contains multiple conserved motifs including SAM domains, ARM/HEATmotifs and a TIR domain (FIG. 1) that mediate oligomerization andprotein-protein interactions (O'Neill, L. A. & Bowie, A. G., Nat. Rev.Immunol., 2007, 7, 353-364; Tewari, R., et al., Trends Cell Biol., 2010,20, 470-481; Qiao, F. & Bowie, J. U., Sci. STKE 2005, re7, 2005). TIRdomains are commonly found in signaling proteins functioning in innateimmunity pathways where they serve as scaffolds for protein complexes(O'Neill, L. A. & Bowie, A. G., Nat. Rev. Immunol., 2007, 7, 353-364).Interestingly, dimerization of SARM1-TIR domains is sufficient to induceaxonal degeneration and to rapidly trigger degradation of NAD+ by actingas the NAD+ cleaving enzyme (Milbrandt et al., WO 2018/057989; Gerdts,J., et al., Science, 2015, 348, 453-457). Given the central role ofSARM1 in the axonal-degeneration pathway and its identified NADaseactivity, efforts have been undertaken to identify agents that canregulate SARM1, and potentially act as useful therapeutic agents, forexample, to protect against neurodegenerative diseases includingperipheral neuropathy, traumatic brain injury, and/or neurodegenerativediseases.

Among other things, the present disclosure provides certain compoundsand/or compositions that act as SARM1 inhibitory agents (e.g., as SARM1inhibitory agents), and technologies relating thereto.

Compounds

In some embodiments, the present disclosure provides a compound ofFormula I:

or a pharmaceutically acceptable salt thereof, wherein:

-   X is N or C—R^(x);-   R^(x) is selected from the group consisting of hydrogen, halogen,    and optionally substituted C₁₋₆ aliphatic;-   R^(a) is selected from the group consisting of hydrogen, halogen,    CN, CF₃, C₁₋₃ aliphatic, and —C(O)N(R^(a)*)₂;-   R^(a)* is selected from hydrogen and C₁₋₃ aliphatic;-   R¹ is selected from the group consisting of hydrogen, halogen,    N(R)₂, OR, C₁₋₆ aliphatic, a 3- to 7-membered monocyclic    heterocyclic ring having 1-2 heteroatoms independently selected from    nitrogen, oxygen, and sulfur, a 5- to 6-membered heteroaryl ring    having 1-3 heteroatoms independently selected from nitrogen, oxygen,    and sulfur, and a 7- to 8-membered bridged bicyclic heterocyclic    ring having 1-2 heteroatoms independently selected from nitrogen,    oxygen, and sulfur, wherein each of C₁₋₆ aliphatic, monocyclic or    bridged bicyclic heterocyclic ring, and heteroaryl ring is    optionally substituted with 1-3 R¹* groups;-   R¹* is selected from oxo, halogen, N(R)₂, OR, C(O)R, S(O)₂R, C(O)OR,    C(O)N(R)₂, optionally substituted C₁₋₆ aliphatic, and an optionally    substituted 5- to 6-membered heteroaryl ring having 1-3 heteroatoms    independently selected from nitrogen, oxygen, and sulfur;-   R is selected from the group consisting of hydrogen, optionally    substituted C₁₋₆ aliphatic, and an optionally substituted 3- to    7-membered saturated monocyclic ring having 0-2 heteroatoms    independently selected from nitrogen, oxygen, and sulfur, or:    -   two R groups, together with the nitrogen atom to which they are        attached, form an optionally substituted 3- to 6-membered        heterocyclic ring containing 0-1 additional heteroatom selected        from nitrogen, oxygen, or sulfur;-   R² is selected from the group consisting of hydrogen, halogen,    S(O)₂R, C(O)OR, C(O)N(R)₂, and an optionally substituted group    selected from C₁₋₆ aliphatic, a 3- to 7-membered heterocyclic ring    having 1-2 heteroatoms independently selected from nitrogen, oxygen,    and sulfur, and a 5- to 6-membered heteroaryl ring having 1-3    heteroatoms independently selected from nitrogen, oxygen, and    sulfur; and-   R³ is selected from the group consisting of hydrogen, halogen, CN,    CF₃, C₁₋₃ aliphatic, and —C(O)N(R^(a)*)₂.

As defined generally above, R^(a) is selected from the group consistingof hydrogen, halogen, —CN, —CF₃, C₁₋₃ aliphatic, and —C(O)N(R^(a)*)₂. Insome embodiments, R^(a) is hydrogen. In some embodiments, R^(a) isselected from the group consisting of halogen, —CN, —CF₃, C₁₋₃aliphatic, and —C(O)N(R^(a)*)₂. In some embodiments, R^(a) is halogen.In some such embodiments, R^(a) is chloro or fluoro.

In some embodiments, R^(a) is —CF₃.

In some embodiments, R^(a) is —CN.

In some embodiments, R^(a) is C₁₋₃ aliphatic. In some such embodiments,R^(a) is —CH₃, —CH₂CH₃, —CH(CH₃)₂, or cyclopropyl.

In some embodiments, R^(a) is —C(O)N(R^(a)*)₂.

As defined generally above, R^(a)* is selected from hydrogen and C₁₋₃aliphatic. In some embodiments, R^(a)* is hydrogen. In some embodiments,R^(a)* is C₁₋₃ aliphatic. In some such embodiments, R^(a)* is —CH₃,—CH₂CH₃, —CH(CH₃)₂, or cyclopropyl.

In some embodiments, R^(a) is —C(O)NH₂. In some embodiments, R^(a) is—C(O)NHCH₃. In some embodiments, R^(a) is —C(O)N(CH₃)₂.

As defined generally above, X is N or C—R^(x). In some embodiments, X isN. In some embodiments, X is C—R^(x). In some embodiments, X is N. Insome embodiments, X is C—R^(x). Accordingly, in some embodiments, thepresent disclosure provides a compound of Formula I-a or I-b:

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R²,R³, and R^(x) is as defined above and as described in classes andsubclasses herein, infra.

As defined generally above, R^(x) is selected from the group consistingof hydrogen, halogen, and optionally substituted C₁₋₆ aliphatic. In someembodiments, R^(x) is hydrogen. In some embodiments, R^(x) is selectedfrom halogen and optionally substituted C₁₋₆ aliphatic. In someembodiments, R^(x) is halogen. In some such embodiments, R^(x) isfluoro, chloro, or bromo.

In some embodiments, the present disclosure provides a compound ofFormula I-C:

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R²,and R³ is as defined above and as described in classes and subclassesherein, infra.

As defined generally above, R¹ is selected from the group consisting ofhydrogen, halogen, N(R)₂, OR, C₁₋₆ aliphatic, a 3- to 7-memberedmonocyclic heterocyclic ring having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur, a 5- to 6-memberedheteroaryl ring having 1-3 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, and a 7- to 8-membered bridged bicyclicheterocyclic ring having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, wherein each of C₁₋₆ aliphatic, monocyclicor bridged bicyclic heterocyclic ring, and heteroaryl ring is optionallysubstituted with 1-3 R¹* groups. In some embodiments, R¹ is hydrogen. Insome embodiments, R¹ is halogen, N(R)₂, OR, C₁₋₆ aliphatic, a 3- to7-membered monocyclic heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen, and sulfur, a 5- to6-membered heteroaryl ring having 1-3 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur, and a 7- to 8-membered bridgedbicyclic heterocyclic ring having 1-2 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur, wherein each of C₁₋₆ aliphatic,monocyclic or bridged bicyclic heterocyclic ring, and heteroaryl ring isoptionally substituted with 1-3 R¹* groups.

In some embodiments, R¹ is halogen. In some such embodiments, R¹ ischloro.

In some embodiments, R¹ is N(R)₂. In some embodiments, R¹ is N(R)₂,wherein R is selected form hydrogen and optionally substituted C₁₋₆aliphatic. In some embodiments, R¹ is NH₂. In some embodiments, R¹ isN(CH₃)₂. In some such embodiments, R¹ is NHR, wherein R is optionallysubstituted C₁₋₆ aliphatic.

In some such embodiments, R¹ is selected from:

In some embodiments, R¹ is N(R)₂, wherein R is selected from hydrogenand an optionally substituted 3- to 7-membered saturated monocyclic ringhaving 0-2 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In some such embodiments, R¹ is NHR, wherein R is an optionallysubstituted 3- to 7-membered saturated monocyclic ring having 0-2heteroatoms independently selected from nitrogen, oxygen, and sulfur. Insome embodiments, R¹ is NHR, wherein R is an optionally substituted 5-to 6-membered saturated monocyclic ring having 0-2 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, R¹ is NHR, wherein R is an optionally substituted 5- to6-membered saturated monocyclic ring having 1 heteroatom selected fromnitrogen, oxygen, and sulfur.

In some embodiments, R¹ is selected from:

In some embodiments, R¹ is OR. In some embodiments, R¹ is OR, wherein Ris optionally substituted C₁₋₆ aliphatic. In some such embodiments, R¹is OCH₃.

In some embodiments, R¹ is C₁₋₆ aliphatic optionally substituted with1-3 R¹* groups. In some such embodiments, R¹ is cyclopropyl.

In some embodiments, R¹ is a 3- to 7-membered monocyclic heterocyclicring having 1-2 heteroatoms independently selected from nitrogen,oxygen, and sulfur, wherein the heterocyclic ring is optionallysubstituted with 1-3 R¹* groups. In some embodiments, R¹ is a 4-memberedmonocyclic heterocyclic ring having 1 heteroatom independently selectedfrom nitrogen, oxygen, and sulfur, wherein the heterocyclic ring isoptionally substituted with 1 R¹* group. In some embodiments, R¹ is

In some embodiments, R¹ is a 5-membered monocyclic heterocyclic ringhaving 1-2 heteroatoms independently selected from nitrogen, oxygen, andsulfur, wherein the heterocyclic ring is optionally substituted with 1-3R¹* groups. In some embodiments, R¹ is a 5-membered monocyclicheterocyclic ring having 1 heteroatom selected from nitrogen, oxygen,and sulfur, wherein the heterocyclic ring is optionally substituted with1-2 R¹* groups. In some embodiments, R¹ is a 5-membered monocyclicheterocyclic ring having 1 nitrogen atom, wherein the heterocyclic ringis optionally substituted with 1 R¹* group. In some embodiments, R¹ isselected from

In some embodiments, R¹ is a 6-membered monocyclic heterocyclic ringhaving 1-2 heteroatoms independently selected from nitrogen, oxygen, andsulfur, wherein the heterocyclic ring is optionally substituted with 1-3R¹* groups. In some embodiments, R¹ is a 6-membered monocyclicheterocyclic ring having 1 heteroatom selected from nitrogen, oxygen,and sulfur, wherein the heterocyclic ring is optionally substituted with1-2 R¹* groups. In some embodiments, R¹ is a 6-membered monocyclicheterocyclic ring having 2 heteroatoms selected from nitrogen, oxygen,and sulfur, wherein the heterocyclic ring is optionally substituted with1-2 R¹* groups.

In some embodiments, R¹ is selected from

In some embodiments, R¹ is a 5- to 6-membered heteroaryl ring having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur,wherein the heteroaryl ring is optionally substituted with 1-3 R¹*groups. In some embodiments, R¹ is a 5-membered heteroaryl ring having1-3 heteroatoms independently selected from nitrogen, oxygen, andsulfur, wherein the heteroaryl ring is optionally substituted with 1-3R¹* groups. In some embodiments, R¹ is a 5-membered heteroaryl ringhaving 1-3 heteroatoms independently selected from nitrogen, oxygen, andsulfur, wherein the heteroaryl ring is optionally substituted with 1-2R¹* groups. In some embodiments R¹ is selected from

In some embodiments, R¹ is a 6-membered heteroaryl ring having 1-2nitrogen atoms, wherein the heteroaryl ring is optionally substitutedwith 1-3 R¹* groups. In some embodiments, R¹ is

In some embodiments, R¹ is a 7- to 8-membered bridged bicyclicheterocyclic ring having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, wherein the bridged bicyclic heterocyclicring is optionally substituted with 1-3 R¹* groups. In some embodiments,R¹ is a 7- to 8-membered bridged bicyclic heterocyclic ring having 1-2nitrogen atoms, wherein the bridged bicyclic heterocyclic ring isoptionally substituted with 1-2 R¹* groups.

In some embodiments, R¹ is selected from

As defined generally above, R¹* is selected from oxo, halogen, N(R)₂,OR, C(O)R, S(O)₂R, C(O)OR, C(O)N(R)₂, optionally substituted C₁₋₆aliphatic, and an optionally substituted 5- to 6-membered heteroarylring having 1-3 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In some embodiments, R¹* is oxo. In someembodiments, R¹* is optionally substituted C₁₋₆ aliphatic. In some suchembodiments, R¹* is —CH₃ or —CH₂CH₃. In some embodiments, R¹* is C₁₋₆aliphatic optionally substituted with —(CH₂)₀₋₄OR^(∘). In some suchembodiments, R^(∘) hydrogen. Accordingly, in some such embodiments, R¹*is C₁₋₆ aliphatic optionally substituted with —(CH₂)₀₋₄OH. In someembodiments, R¹* is —CH₂OH.

In some embodiments, R¹* is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄C(O)N(R^(∘))₂. In some such embodiments, R^(∘) hydrogen.Accordingly, in some such embodiments, R¹* is C₁₋₆ aliphatic optionallysubstituted with —(CH₂)₀₋₄C(O)NH₂. In some embodiments, R¹* is—CH₂CH₂C(O)NH₂.

In some embodiments, R¹* is N(R)₂. In some such embodiments, R isselected from hydrogen and optionally substituted C₁₋₆ aliphatic. Insome embodiments, R¹* is NH₂. In some embodiments, R¹* is NHR, wherein Ris optionally substituted C₁₋₆ aliphatic. In some embodiments, R¹* isNHR, wherein R is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄OR^(∘). In some such embodiments, R^(∘) hydrogen. Accordingly,in some such embodiments, R¹* is NHR, wherein R is C₁₋₆ aliphaticoptionally substituted with —(CH₂)₀₋₄OH. In some embodiments, R¹* is—NHCH₂CH₂OH.

In some embodiments, R¹* is —OR. In some such embodiments, R is hydrogenor optionally substituted C₁₋₆ aliphatic. In some embodiments, R¹* is—OH or —OCH₃.

In some embodiments, R¹* is C(O)N(R)₂. In some such embodiments, each Ris selected from hydrogen and optionally substituted C₁₋₆ aliphatic. Insome embodiments, R¹* is selected from —C(O)NH₂, —C(O)NHCH₃, andC(O)N(CH₃)₂.

In some embodiments, R¹* is C(O)R. In some such embodiments, R isoptionally substituted C₁₋₆ aliphatic. In some embodiments, R¹* is—C(O)CH₃.

In some embodiments, R¹* is S(O)₂R. In some such embodiments, R isoptionally substituted C₁₋₆ aliphatic. In some embodiments, R¹* is—S(O)₂CH₃.

In some embodiments, R¹* is an optionally substituted 5- to 6-memberedheteroaryl ring having 1-3 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In some embodiments, R¹* is an optionallysubstituted 5-membered heteroaryl ring having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, R¹* is an optionally substituted 5-membered heteroaryl ringhaving 2 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In some embodiments, R¹* is

In some embodiments, R¹* is selected from oxo, —CH₃, —CH₂CH₃, —CH₂OH,—CH₂CH₂C(O)NH₂, —OH, —OCH₃, —NH₂, —NHCH₂CH₂OH, —C(O)NH₂, —C(O)NHCH₃,C(O)N(CH₃)₂, —C(O)CH₃, —S(O)₂CH₃, and

As defined generally above, R is selected from the group consisting ofhydrogen, optionally substituted C₁₋₆ aliphatic, and an optionallysubstituted 3- to 7-membered saturated monocyclic ring having 0-2heteroatoms independently selected from nitrogen, oxygen, and sulfur,or: two R groups, together with the nitrogen atom to which they areattached, form an optionally substituted 3- to 6-membered heterocyclicring containing 0-1 additional heteroatom selected from nitrogen,oxygen, or sulfur. In some embodiments, R is hydrogen. In someembodiments, R is optionally substituted C₁₋₆ aliphatic or an optionallysubstituted 3- to 7-membered saturated monocyclic ring having 0-2heteroatoms independently selected from nitrogen, oxygen, and sulfur.or: two R groups, together with the nitrogen atom to which they areattached, form an optionally substituted 3- to 6-membered heterocyclicring containing 0-1 additional heteroatom selected from nitrogen,oxygen, or sulfur.

In some embodiments, R is hydrogen or C₁₋₆ aliphatic. In some suchembodiments, R is hydrogen or —CH₃. In some embodiments, R is hydrogenor optionally substituted C₁₋₆ aliphatic.

In some embodiments, R is C₁₋₆ aliphatic. In some embodiments, R is—CH₃. In some embodiments, R is cyclopropyl.

In some embodiments, R is C₁₋₆ aliphatic optionally substituted with agroup selected from —(CH₂)₀₋₄R^(∘), —(CH₂)₀₋₄OR^(∘),—(CH₂)₀₋₄C(O)N(R^(∘))₂, and —(CH₂)₀₋₄N(R^(∘))C(O)N(R^(∘))₂. In some suchembodiments, R^(∘) is selected from hydrogen, C₁₋₆ aliphatic, a 5- to6-membered saturated, partially unsaturated, or aryl ring having 0-4heteroatoms independently selected from nitrogen, oxygen, or sulfur, oran 8- to 10-membered bicyclic aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur. In someembodiments, R^(∘) is substituted with a group selected from—(CH₂)₀₋₂R^(●), —(CH₂)₀₋₂OH, —(CH₂)₀₋₂OR^(●), and —(CH₂)₀₋₂C(O)NH₂,wherein R^(●) is C₁₋₄ aliphatic.

In some embodiments, R is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄R^(∘), wherein R^(∘) is C₁₋₆ aliphatic, a 5- to 6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or an 8- to10-membered bicyclic aryl ring having 0-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur.

In some embodiments, R is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄R^(∘), wherein R^(∘) is a 5- to 6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. In some embodiments, R is C₁₋₆aliphatic optionally substituted with —(CH₂)₀₋₄R^(∘), wherein R^(∘) is a5-membered heteroaryl ring having 1-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. In some embodiments, R is C₁₋₆aliphatic optionally substituted with —(CH₂)₀₋₄R^(∘), wherein R^(∘) is a5-membered heteroaryl ring having 1-2 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. In some embodiments, R is C₁₋₆aliphatic optionally substituted with —(CH₂)₀₋₄R^(∘), wherein R^(∘) is a5-membered heteroaryl ring having 1-2 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur, wherein R^(∘) is substituted with—(CH₂)₀₋₂R^(●). In some embodiments, R is C₁₋₆ aliphatic optionallysubstituted with

In some embodiments, R is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄R^(∘), wherein R^(∘) is a 6-membered heteroaryl ring having 1-2nitrogen atoms. In some embodiments, R is C₁₋₆ aliphatic optionallysubstituted with

In some embodiments, R is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄R^(∘), wherein R^(∘) is a 5- to 6-membered saturated ringhaving 1-2 heteroatoms independently selected from nitrogen, oxygen, orsulfur. In some embodiments, R is C₁₋₆ aliphatic optionally substitutedwith —(CH₂)₀₋₄R^(∘), wherein R^(∘) is a 6-membered saturated ring having1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur.In some embodiments, R is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄R^(∘), wherein R^(∘) is a 6-membered saturated ring having 1-2heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein R^(∘) is substituted with —(CH₂)₀₋₂R^(●). In some embodiments, Ris C₁₋₆ aliphatic optionally substituted with

In some embodiments, R is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄R^(∘), wherein R^(∘) is phenyl. In some such embodiments, R^(∘)is substituted with —(CH₂)₀₋₂C(O)NH₂. In some embodiments, R is C₁₋₆aliphatic optionally substituted with

In some embodiments, R is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄R^(∘), wherein R^(∘) is an 8- to 10-membered bicyclic aryl ringhaving 0-4 heteroatoms independently selected from nitrogen, oxygen, orsulfur. In some embodiments, R is C₁₋₆ aliphatic optionally substitutedwith —(CH₂)₀₋₄R^(∘), wherein R^(∘) is a 9-membered bicyclic aryl ringhaving 1-3 heteroatoms independently selected from nitrogen, oxygen, orsulfur. In some such embodiments, R^(∘) is substituted with—(CH₂)₀₋₂R^(●). In some embodiments, R is C₁₋₆ aliphatic optionallysubstituted with

In some embodiments, R is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄OR^(∘), wherein R^(∘) is hydrogen or C₁₋₆ aliphatic. In someembodiments, R is C₁₋₆ aliphatic optionally substituted with —OR^(∘),wherein R^(∘) is hydrogen or C₁₋₆ aliphatic. In some embodiments, R isC₁₋₆ aliphatic optionally substituted with —OH.

In some embodiments, R is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄C(O)N(R^(∘))₂. In some embodiments, R is C₁₋₆ aliphaticoptionally substituted with —(CH₂)₀₋₄C(O)N(R^(∘))₂, wherein R^(∘) ishydrogen or C₁₋₆ aliphatic. In some embodiments, R is C₁₋₆ aliphaticoptionally substituted with —(CH₂)₀₋₄C(O)N(R^(∘))₂, wherein R^(∘) ishydrogen or C₁₋₆ aliphatic optionally substituted with —(CH₂)₀₋₂OR^(●).In some embodiments, R is C₁₋₆ aliphatic optionally substituted with—C(O)N(R^(∘))₂, wherein R^(∘) is hydrogen or C₁₋₆ aliphatic optionallysubstituted with —(CH₂)₀₋₂OR^(●). In some embodiments, R is C₁₋₆aliphatic optionally substituted with —C(O)NHCH₂CH₂OCH₃. In someembodiments, R is C₁₋₆ aliphatic optionally substituted with —C(O)NHCH₃.

In some embodiments, R is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄N(R^(∘))C(O)N(R^(∘))₂. In some embodiments, R is C₁₋₆ aliphaticoptionally substituted with —(CH₂)₀₋₄N(R^(∘))C(O)N(R^(∘))₂, whereinR^(∘) is hydrogen or C₁₋₆ aliphatic. In some embodiments, R is C₁₋₆aliphatic optionally substituted with —NHC(O)N(CH₃)₂.

In some embodiments, R is C₁₋₆ aliphatic optionally substituted with —OHand a 5- to 6-membered saturated, partially unsaturated, or aryl ringhaving 0-4 heteroatoms independently selected from nitrogen, oxygen, orsulfur. In some embodiments, R is C₁₋₆ aliphatic optionally substitutedwith —OH and —(CH₂)₀₋₄C(O)N(R^(∘))₂.

In some embodiments, R is an optionally substituted 3- to 7-memberedsaturated monocyclic ring having 0-2 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur. In some embodiments, R is a7-membered saturated monocyclic ring having 1 heteroatom selected fromnitrogen, oxygen, and sulfur. In some such embodiments, R is substitutedon a carbon atom with ═O. In some embodiments, R is

In some embodiments, R is an optionally substituted 3- to 7-memberedsaturated monocyclic ring having 0-2 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur. In some embodiments, R is a6-membered saturated monocyclic ring having 1 heteroatom selected fromnitrogen, oxygen, and sulfur. In some such embodiments, R is substitutedwith —(CH₂)₀₋₄C(O)NR^(∘) ₂. In some embodiments, R is a 6-memberedsaturated monocyclic ring having 1 heteroatom selected from nitrogen,oxygen, and sulfur, substituted with —C(O)NH₂. In some embodiments, R is

In some embodiments, R is a 6-membered saturated monocyclic ring having1 heteroatom selected from nitrogen, oxygen, and sulfur, wherein R issubstituted with —(CH₂)₀₋₄R^(∘). In some such embodiments, R^(∘) is C₁₋₆aliphatic substituted with —(CH₂)₀₋₂R^(●), wherein R^(●) is a 3- to6-membered saturated, partially unsaturated, or aryl ring having 0-4heteroatoms independently selected from nitrogen, oxygen, or sulfur. Insome embodiments, R is

In some embodiments, R is a 5-membered saturated monocyclic ring having1 heteroatom selected from nitrogen, oxygen, and sulfur. In some suchembodiments, R is substituted with ═O and —(CH₂)₀₋₄R^(∘). In someembodiments, R is

In some embodiments, R is selected from hydrogen, C₁₋₆ aliphaticoptionally substituted with one or more groups selected from —OH,—NHC(O)N(CH₃)₂, —C(O)NHCH₃, —C(O)NHCH₂CH₂OCH₃,

or R is

In some embodiments, R¹ is selected from the group consisting ofhydrogen, halogen, —NH₂, —OCH₃,

As defined generally above, R² is selected from the group consisting ofhydrogen, halogen, S(O)₂R, C(O)OR, C(O)N(R)₂, and an optionallysubstituted group selected from C₁₋₆ aliphatic, a 3- to 7-memberedheterocyclic ring having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, and a 5- to 6-membered heteroaryl ringhaving 1-3 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In some embodiments, R² is hydrogen. In some embodiments, R² isselected from the group consisting of halogen, S(O)₂R, C(O)OR,C(O)N(R)₂, and an optionally substituted group selected from C₁₋₆aliphatic, a 3- to 7-membered heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen, and sulfur, and a 5- to6-membered heteroaryl ring having 1-3 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur.

In some embodiments, R² is halogen. In some such embodiments, R² isfluoro, chloro or bromo.

In some embodiments, R² is optionally substituted C₁₋₆ aliphatic. Insome such embodiments, R² is —CH₃ or —CH₂CH₃. In some embodiments, R² iscyclopropyl. In some embodiments, R² is C₁₋₆ aliphatic optionallysubstituted with —(CH₂)₀₋₄OR^(∘). In some such embodiments, R^(∘) ishydrogen. Accordingly, in some embodiments, R² is C₁₋₆ aliphaticoptionally substituted with —(CH₂)₀₋₄OH. In some embodiments, R² is—C(OH)(CH₃)₂.

In some embodiments, R² is C(O)OR. In some such embodiments, R ishydrogen. Accordingly, in some embodiments, R² is C(O)OH.

In some embodiments, R² is S(O)₂R. In some such embodiments, R isoptionally substituted C₁₋₆ aliphatic. In some embodiments, R² is—S(O)₂CH₃.

In some embodiments, R² is C(O)N(R)₂. In some such embodiments, each Ris selected from hydrogen and optionally substituted C₁₋₆ aliphatic. Insome embodiments, R² is C(O)NH₂. In some embodiments, R² is C(O)NHR,wherein R is optionally substituted C₁₋₆ aliphatic. In some embodiments,R² is C(O)NHR, wherein R is C₁₋₆ aliphatic optionally substituted with—(CH₂)₀₋₄OR^(∘). In some embodiments, R² is C(O)NHR, wherein R is C₁₋₆aliphatic optionally substituted with —(CH₂)₀₋₄OR^(∘) and R^(∘) ishydrogen. In some embodiments, R² is C(O)NHR, wherein R is C₁₋₆aliphatic optionally substituted with —(CH₂)₀₋₄OH. In some embodiments,R² is —C(O)NHCH₂CH₂OH.

In some embodiments, R² is C(O)N(R)₂, wherein two R groups, togetherwith the nitrogen atom to which they are attached, form an optionallysubstituted 3- to 6-membered heterocyclic ring containing 0-1 additionalheteroatom selected from nitrogen, oxygen, or sulfur. In someembodiments, R² is C(O)N(R)₂, wherein two R groups, together with thenitrogen atom to which they are attached, form an optionally substituted6-membered heterocyclic ring containing 0-1 additional heteroatomselected from nitrogen, oxygen, or sulfur. In some embodiments, R² is

In some embodiments, R² is an optionally substituted 3- to 7-memberedheterocyclic ring having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In some embodiments, R² is a 4- to6-membered heterocyclic ring having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur. In some embodiments, R² is a4-membered heterocyclic ring having 1 heteroatom selected from nitrogen,oxygen, and sulfur. In some such embodiments, R² is oxetanyl. In someembodiments, R² is a 5-membered heterocyclic ring having 1 heteroatomselected from nitrogen, oxygen, and sulfur. In some such embodiments, R²is tetrahydrofuranyl. In some embodiments, R² is a 6-memberedheterocyclic ring having 1 heteroatom selected from nitrogen, oxygen,and sulfur. In some such embodiments, R² is morpholinyl.

In some embodiments, R² is an optionally substituted 5- to 6-memberedheteroaryl ring having 1-3 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In some embodiments, R² is a 5-memberedheteroaryl ring having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In some embodiments, R² is pyrazolyl. Insome such embodiments, R² is 3-methyl-1H-pyrazol-4-yl. In someembodiments, R² is 1-methyl-1H-pyrazol-4-yl. In some embodiments, R² is1H-pyrazol-4-yl.

In some embodiments, R² is an optionally substituted 6-memberedheteroaryl ring having 1-3 nitrogen atoms. In some embodiments, R² ispyridin-3-yl. In some embodiments, R² is 2-methyl-pyridin-3-yl.

In some embodiments, R² is selected from the group consisting ofhydrogen, halogen, —CH₃, CH₂CH₃, cyclopropyl, —C(OH)(CH₃)₂, —C(O)OH,—S(O)₂CH₃, —C(O)NH₂, —C(O)NHCH₃, —C(O)N(CH₃)₂, —C(O)NHCH₂CH₂OH, or agroup selected from

As defined generally above, R³ is selected from the group consisting ofhydrogen, halogen, CN, CF₃, C₁₋₃ aliphatic, and —C(O)N(R^(a)*)₂. In someembodiments, R³ is hydrogen. In some embodiments, R³ is halogen, CN,CF₃, C₁₋₃ aliphatic, and —C(O)N(R^(a)*)₂. In some embodiments, R³ ishalogen. In some such embodiments, R³ is fluoro or chloro.

In some embodiments, R³ is CN. In some embodiments, R³ is CF₃. In someembodiments, R³ is C₁₋₃ aliphatic. In some such embodiments, R³ is —CH₃,—CH₂CH₃, —CH(CH₃)₂, or cyclopropyl. In some embodiments, R³ is—C(O)N(R^(a)*)₂. In some embodiments, R³ is —C(O)NH₂. In someembodiments, R³ is —C(O)NHCH₃. In some embodiments, R³ is —C(O)N(CH₃)₂.

In some embodiments of Formulae I-a, I-b, and I-c, R³ is hydrogen.Accordingly, in some embodiments, the present disclosure provides acompound of Formula I-a-i, I-b-i, and I-c-i:

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R²,and R^(x) is as defined above and described in classes and subclassesherein.

In some embodiments of Formulae I-a-i, I-b-i, and I-c-i, R² is chloro.Accordingly, in some embodiments, the present disclosure provides acompound of Formula I-a-ii, I-b-ii, and

or a pharmaceutically acceptable salt thereof, wherein each of R¹ andR^(x) is as defined above and described in classes and subclassesherein.

In some embodiments, a compound of Formula I is selected from:

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In some embodiments, the present disclosure provides compounds having astructure as set forth in Formula II:

or a pharmaceutically acceptable salt thereof, wherein:

-   X is Nor C—R^(x);-   R^(x) is selected from the group consisting of hydrogen, halogen,    and optionally substituted C₁₋₆ aliphatic;-   R¹ is selected from the group consisting of a 3- to 7-membered    monocyclic heterocyclic ring having 1-2 heteroatoms independently    selected from nitrogen, oxygen, and sulfur, a 5- to 6-membered    heteroaryl ring having 1-3 heteroatoms independently selected from    nitrogen, oxygen, and sulfur, and a 7- to 8-membered bridged    bicyclic heterocyclic ring having 1-2 heteroatoms independently    selected from nitrogen, oxygen, and sulfur, wherein each of C₁₋₆    aliphatic, monocyclic or bridged bicyclic heterocyclic ring, and    heteroaryl ring is optionally substituted with 1-3 R¹* groups;-   R¹* is selected from oxo, halogen, N(R)₂, OR, C(O)R, S(O)₂R, C(O)OR,    C(O)N(R)₂, and C₁₋₆ aliphatic;-   R is selected from the group consisting of hydrogen and optionally    substituted C₁₋₆ aliphatic, or:    -   two R groups, together with the nitrogen atom to which they are        attached, form an optionally substituted 3- to 6-membered        heterocyclic ring containing 0-1 additional heteroatom selected        from nitrogen, oxygen, or sulfur;-   R² is selected from the group consisting of hydrogen, halogen,    S(O)₂R, C(O)OR, C(O)N(R)₂, and an optionally substituted group    selected from C₁₋₆ aliphatic, a 3- to 7-membered heterocyclic ring    having 1-2 heteroatoms independently selected from nitrogen, oxygen,    and sulfur, and a 5- to 6-membered heteroaryl ring having 1-3    heteroatoms independently selected from nitrogen, oxygen, and    sulfur; and-   R³ is selected from the group consisting of hydrogen, halogen, CN,    CF₃, C₁₋₃ aliphatic, and —C(O)N(R^(a)*)₂.

In some embodiments, the present disclosure provides compounds having astructure as set forth in Formulae II-a, II-b, and II-c:

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R²,R³, and R^(x) is as defined above for Formula II and as described inclasses herein, supra.

Enumerated Embodiments

Embodiment 1. A compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein:

-   X is N or C—R^(x);-   R^(x) is selected from the group consisting of hydrogen, halogen,    and optionally substituted C₁₋₆ aliphatic;-   R^(a) is selected from the group consisting of hydrogen, halogen,    CN, CF₃, C₁₋₃ aliphatic, and —C(O)N(R^(a)*)₂.-   R^(a)* is selected from hydrogen and C₁₋₃ aliphatic;-   R¹ is selected from the group consisting of hydrogen, halogen,    N(R)₂, OR, C₁₋₆ aliphatic, a 3- to 7-membered monocyclic    heterocyclic ring having 1-2 heteroatoms independently selected from    nitrogen, oxygen, and sulfur, a 5- to 6-membered heteroaryl ring    having 1-3 heteroatoms independently selected from nitrogen, oxygen,    and sulfur, and a 7- to 8-membered bridged bicyclic heterocyclic    ring having 1-2 heteroatoms independently selected from nitrogen,    oxygen, and sulfur, wherein each of C₁₋₆ aliphatic, monocyclic or    bridged bicyclic heterocyclic ring, and heteroaryl ring is    optionally substituted with 1-3 R¹* groups;-   R¹* is selected from oxo, halogen, N(R)₂, OR, C(O)R, S(O)₂R, C(O)OR,    C(O)N(R)₂, optionally substituted C₁₋₆ aliphatic, and an optionally    substituted 5- to 6-membered heteroaryl ring having 1-3 heteroatoms    independently selected from nitrogen, oxygen, and sulfur;-   R is selected from the group consisting of hydrogen, an optionally    substituted C₁₋₆ aliphatic, and an optionally substituted 3- to    7-membered saturated monocyclic ring having 0-2 heteroatoms    independently selected from nitrogen, oxygen, and sulfur, or:    -   two R groups, together with the nitrogen atom to which they are        attached, form an optionally substituted 3- to 6-membered        heterocyclic ring containing 0-1 additional heteroatom selected        from nitrogen, oxygen, or sulfur;-   R² is selected from the group consisting of hydrogen, halogen,    S(O)₂R, C(O)OR, C(O)N(R)₂, and an optionally substituted group    selected from C₁₋₆ aliphatic, a 3- to 7-membered heterocyclic ring    having 1-2 heteroatoms independently selected from nitrogen, oxygen,    and sulfur, and a 5- to 6-membered heteroaryl ring having 1-3    heteroatoms independently selected from nitrogen, oxygen, and    sulfur; and-   R³ is selected from the group consisting of hydrogen, halogen, CN,    CF₃, C₁₋₃ aliphatic, and —C(O)N(R^(a)*)₂.

Embodiment 2. The compound according to embodiment 1, wherein R^(a) isCN.

Embodiment 3. The compound according to embodiment 2, wherein thecompound is a compound of Formula I-a or I-b:

or a pharmaceutically acceptable salt thereof.

Embodiment 4. The compound according to embodiment 3, wherein R^(x) ishalogen.

Embodiment 5. The compound according to embodiment 3, wherein R^(x) ishydrogen.

Embodiment 6. The compound according to embodiment 5, wherein thecompound is a compound of Formula I-c:

or a pharmaceutically acceptable salt thereof.

Embodiment 7. The compound according to any one of embodiments 1-6,wherein R¹ is a 3- to 7-membered monocyclic heterocyclic ring having 1-2heteroatoms independently selected from nitrogen, oxygen, and sulfur,wherein the heterocyclic ring is optionally substituted with 1-3 R¹*groups.

Embodiment 8. The compound according to embodiment 7, wherein R¹ is a5-membered monocyclic heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen, and sulfur, wherein theheterocyclic ring is optionally substituted with 1-3 R¹* groups.

Embodiment 9. The compound according to embodiment 8, wherein R¹ is a5-membered monocyclic heterocyclic ring having 1 heteroatom selectedfrom nitrogen, oxygen, and sulfur, wherein the heterocyclic ring isoptionally substituted with 1-2 R¹* groups.

Embodiment 10. The compound according to embodiment 9, wherein R¹ is a5-membered monocyclic heterocyclic ring having 1 nitrogen atom, whereinthe heterocyclic ring is optionally substituted with 1 R¹* group.

Embodiment 11. The compound according to embodiment 10, wherein R¹ isselected from

Embodiment 12. The compound according to embodiment 7, wherein, R¹ is a6-membered monocyclic heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen, and sulfur, wherein theheterocyclic ring is optionally substituted with 1-3 R¹* groups.

Embodiment 13. The compound according to embodiment 12, wherein R¹ is a6-membered monocyclic heterocyclic ring having 1 heteroatom selectedfrom nitrogen, oxygen, and sulfur, wherein the heterocyclic ring isoptionally substituted with 1-2 R¹* groups.

Embodiment 14. The compound according to embodiment 13, wherein R¹ is a6-membered monocyclic heterocyclic ring having 2 heteroatoms selectedfrom nitrogen, oxygen, and sulfur, wherein the heterocyclic ring isoptionally substituted with 1-2 R¹* groups.

Embodiment 15. The compound according to embodiment 14, wherein R¹ isselected from

Embodiment 16. The compound according to any one of embodiments 1-6,wherein R¹ is a 5- to 6-membered heteroaryl ring having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur, wherein theheteroaryl ring is optionally substituted with 1-3 R¹* groups.

Embodiment 17. The compound according to embodiment 16, wherein R¹ is a5-membered heteroaryl ring having 1-3 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur, wherein the heteroaryl ring isoptionally substituted with 1-3 R¹* groups.

Embodiment 18. The compound according to embodiment 17, wherein R¹ is a5-membered heteroaryl ring having 1-3 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur, wherein the heteroaryl ring isoptionally substituted with 1-2 R¹* groups.

Embodiment 19. The compound according to embodiment 18, wherein R¹ isselected from

Embodiment 20. The compound according to embodiment 16, wherein R¹ is a6-membered heteroaryl ring having 1-2 nitrogen atoms, wherein theheteroaryl ring is optionally substituted with 1-3 R¹* groups.

Embodiment 21. The compound according to embodiment 20, wherein R¹ is

Embodiment 22. The compound according to any one of embodiments 1-6,wherein R¹ is a 7- to 8-membered bridged bicyclic heterocyclic ringhaving 1-2 heteroatoms independently selected from nitrogen, oxygen, andsulfur, wherein the bridged bicyclic heterocyclic ring is optionallysubstituted with 1-3 R¹* groups.

Embodiment 23. The compound according to embodiment 22, wherein R¹ is a7- to 8-membered bridged bicyclic heterocyclic ring having 1-2 nitrogenatoms, wherein the bridged bicyclic heterocyclic ring is optionallysubstituted with 1-2 R¹* groups.

Embodiment 24. The compound according to embodiment 23, wherein R¹ isselected from

Embodiment 25. The compound according to any one of embodiments 7-24,wherein R¹* is N(R)₂.

Embodiment 26. The compound according to any one of embodiments 7-24,wherein R¹* is C(O)N(R)₂.

Embodiment 27. The compound according to any one of embodiments 7-24,wherein R¹* is OR.

Embodiment 28. The compound according to any one of embodiments 7-24,wherein R¹* is C(O)R.

Embodiment 29. The compound according to any one of embodiments 7-24,wherein R¹* is S(O)₂R.

Embodiment 30. The compound according to embodiments 25 or 26, wherein Ris selected from hydrogen and optionally substituted C₁₋₆ aliphatic.

Embodiment 31. The compound according to embodiment 30, wherein each Ris hydrogen.

Embodiment 32. The compound according to embodiment 30, wherein each Ris —CH₃.

Embodiment 33. The compound according to embodiment 30, wherein one R ishydrogen and one R is optionally substituted C₁₋₆ aliphatic.

Embodiment 34. The compound according to embodiment 27, wherein R ishydrogen.

Embodiment 35. The compound according to any one of embodiments 27-29,wherein R is —CH₃.

Embodiment 36. The compound according to any one of embodiments 7-24,wherein R¹* is C₁₋₆ aliphatic.

Embodiment 37. The compound according to embodiment 36, wherein R¹* is—CH₃.

Embodiment 38. The compound according to any one of embodiments 7-24,wherein R¹ is selected from

Embodiment 39. The compound according to any one of embodiments 1-6,wherein R¹ is N(R)₂.

Embodiment 40. The compound according to embodiment 39, wherein each Ris hydrogen.

Embodiment 41. The compound according to embodiment 39, wherein each Ris —CH₃.

Embodiment 42. The compound according to embodiment 39, wherein R¹ is—NHCH₃.

Embodiment 43. The compound according to embodiment 39, wherein R¹ is

Embodiment 44. The compound according to embodiment 39, wherein R¹ is

Embodiment 45. The compound according to any one of embodiments 1-6,wherein R¹ is —OR.

Embodiment 46. The compound according to embodiment 45, wherein R ishydrogen.

Embodiment 47. The compound according to embodiment 45, wherein R is—CH₃.

Embodiment 48. The compound according to any one of embodiments, 1-47,wherein R² is halogen.

Embodiment 49. The compound according to embodiment 48, wherein R² ischloro.

Embodiment 50. The compound according to embodiment 48, wherein R² isbromo.

Embodiment 51. The compound according to embodiment 48, wherein R² isfluoro.

Embodiment 52. The compound according to any one of embodiments, 1-47,wherein R² is —CH₃.

Embodiment 53. The compound according to any one of embodiments, 1-47,wherein R² is hydrogen.

Embodiment 54. The compound according to any one of embodiments, 1-53,wherein R³ is hydrogen.

Embodiment 54. The compound according to any one of embodiments, 1-53,wherein R³ is halogen.

Embodiment 55. The compound according to embodiment 54, wherein R³ ischloro.

Embodiment 56. The compound according to embodiment 55, wherein thecompound a compound of Formula I-a-ii, I-b-ii, or I-c-ii:

or a pharmaceutically acceptable salt thereof.

Embodiment 57. The compound according to embodiment 1, wherein thecompound is a compound of Formula II:

or a pharmaceutically acceptable salt thereof, wherein:

-   X is N or C—R^(x);-   R^(x) is selected from the group consisting of hydrogen, halogen,    and optionally substituted C₁₋₆ aliphatic;-   R¹ is selected from the group consisting of a 3- to 7-membered    monocyclic heterocyclic ring having 1-2 heteroatoms independently    selected from nitrogen, oxygen, and sulfur, a 5- to 6-membered    heteroaryl ring having 1-3 heteroatoms independently selected from    nitrogen, oxygen, and sulfur, and a 7- to 8-membered bridged    bicyclic heterocyclic ring having 1-2 heteroatoms independently    selected from nitrogen, oxygen, and sulfur, wherein each of C₁₋₆    aliphatic, monocyclic or bridged bicyclic heterocyclic ring, and    heteroaryl ring is optionally substituted with 1-3 R¹* groups;-   R¹* is selected from oxo, halogen, N(R)₂, OR, C(O)R, S(O)₂R, C(O)OR,    C(O)N(R)₂, and C₁₋₆ aliphatic;-   R is selected from the group consisting of hydrogen and optionally    substituted C₁₋₆ aliphatic, or:    -   two R groups, together with the nitrogen atom to which they are        attached, form an optionally substituted 3- to 6-membered        heterocyclic ring containing 0-1 additional heteroatom selected        from nitrogen, oxygen, or sulfur;-   R² is selected from the group consisting of hydrogen, halogen,    S(O)₂R, C(O)OR, C(O)N(R)₂, and an optionally substituted group    selected from C₁₋₆ aliphatic, a 3- to 7-membered heterocyclic ring    having 1-2 heteroatoms independently selected from nitrogen, oxygen,    and sulfur, and a 5- to 6-membered heteroaryl ring having 1-3    heteroatoms independently selected from nitrogen, oxygen, and    sulfur; and-   R³ is selected from the group consisting of hydrogen, halogen, CN,    CF₃, C₁₋₃ aliphatic, and —C(O)N(R^(a)*)₂.

Embodiment 58. The compound according to embodiment 57, wherein thecompound is a compound of Formula II-a, II-b, or II-c:

or a pharmaceutically acceptable salt thereof.

Compositions

In some embodiments, a compound of Formula I or Formula II may beprovided in a composition, e.g., in combination (e.g., admixture) withone or more other components.

In some embodiments, the present disclosure provides compositions thatcomprise and/or deliver a compound of Formula I or Formula II, or anactive metabolite thereof, e.g., when contacted with or otherwiseadministered to a system or environment e.g., which system orenvironment may include SARM1 NADase activity; in some embodiments,administration of such a composition to the system or environmentachieves inhibition of SARM1 activity as described herein.

In some embodiments, a provided composition as described herein may be apharmaceutical composition in that it comprises an active agent and oneor more pharmaceutically acceptable excipients; in some suchembodiments, a provided pharmaceutical composition comprises and/ordelivers a compound of Formula I or Formula II, or an active metabolitethereof to a relevant system or environment (e.g., to a subject in needthereof) as described herein.

In some embodiments, one or more compounds of Formula I or Formula II isprovided and/or utilized in a pharmaceutically acceptable salt form.

Among other things, the present disclosure provides compositionscomprising a compound of Formula I or Formula II, or a pharmaceuticallyacceptable salt or derivative thereof, and a pharmaceutically acceptablecarrier, adjuvant, or vehicle. The amount of compound in providedcompositions is such that is effective to measurably inhibit axonaldegeneration in a biological sample or in a patient. In certainembodiments, a provided compound or composition is formulated foradministration to a patient in need of such composition. The compoundsand compositions, according to the methods of the present disclosure,may be administered using any amount and any route of administrationeffective for treating or lessening the severity of any disease ordisorder described herein. Provided compounds are preferably formulatedin dosage unit form for ease of administration and uniformity of dosage.The expression “dosage unit form” as used herein refers to a physicallydiscrete unit of agent appropriate for the patient to be treated. Itwill be understood, however, that the total daily usage of the providedcompounds and compositions will be decided by the attending physicianwithin the scope of sound medical judgment. The specific effective doselevel for any particular patient or organism will vary from subject tosubject, depending on a variety of factors, including the disorder beingtreated and the severity of the disorder; the activity of the specificcompound employed; the specific composition employed and its route ofadministration; the species, age, body weight, sex and diet of thepatient; the general condition of the subject; the time ofadministration; the rate of excretion of the specific compound employed;the duration of the treatment; drugs used in combination or coincidentalwith the specific compound employed, and the like.

Provided compositions may be administered orally, parenterally, byinhalation or nasal spray, topically (e.g., as by powders, ointments, ordrops), rectally, buccally, intravaginally, intraperitoneally,intracisternally or via an implanted reservoir, depending on theseverity of the condition being treated. Preferably, the compositionsare administered orally, intraperitoneally or intravenously. In certainembodiments, provided compounds are administered orally or parenterallyat dosage levels of about 0.01 mg/kg to about 50 mg/kg, of subject bodyweight per day, one or more times a day, to obtain the desiredtherapeutic effect.

The term “parenteral” as used herein includes subcutaneous, intravenous,intramuscular, intra-articular, intra-synovial, intrasternal,intrathecal, intrahepatic, intralesional and intracranial injection orinfusion techniques. Sterile injectable forms of provided compositionsmay be aqueous or oleaginous suspension. These suspensions may beformulated according to techniques known in the art using suitabledispersing or wetting agents and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a nontoxic parenterally acceptable diluent or solvent, forexample, as a solution in 1,3-butanediol. Among the acceptable vehiclesand solvents that may be employed are water, Ringer's solution andisotonic sodium chloride solution. In addition, sterile, fixed oils areconventionally employed as a solvent or suspending medium.

For this purpose, any bland fixed oil may be employed includingsynthetic mono- or di-glycerides. Fatty acids, such as oleic acid andits glyceride derivatives are useful in the preparation of injectables,as are natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant, such as carboxymethyl cellulose or similar dispersingagents that are commonly used in the formulation of pharmaceuticallyacceptable dosage forms including emulsions and suspensions. Othercommonly used surfactants, such as Tweens, Spans and other emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms may also be used for the purposes of formulation.

Injectable formulations can be sterilized, for example, by filtrationthrough a bacterial-retaining filter, or by incorporating sterilizingagents in the form of sterile solid compositions which can be dissolvedor dispersed in sterile water or other sterile injectable medium priorto use.

In order to prolong the effect of a provided compound, it is oftendesirable to slow the absorption of the compound from subcutaneous orintramuscular injection. This may be accomplished by the use of a liquidsuspension of crystalline or amorphous material with poor watersolubility. The rate of absorption of the compound then depends upon itsrate of dissolution that, in turn, may depend upon crystal size andcrystalline form. Alternatively, delayed absorption of a parenterallyadministered compound form is accomplished by dissolving or suspendingthe compound in an oil vehicle. Injectable depot forms are made byforming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Pharmaceutically acceptable compositions described herein may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In suchsolid dosage forms the active compound may be admixed with at least oneinert diluent such as sucrose, lactose or starch. Such dosage forms mayalso comprise, as is normal practice, additional substances other thaninert diluents, e.g., lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. When aqueoussuspensions are required for oral use, the active ingredient is combinedwith emulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and/or i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents. The active compounds canalso be in micro-encapsulated form with one or more excipients as notedabove.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings (i.e. buffering agents) and other coatings well known in thepharmaceutical formulating art. They may optionally contain opacifyingagents and can also be of a composition that they release the activeingredient(s) only, or preferentially, in a certain part of theintestinal tract, optionally, in a delayed manner. Examples of embeddingcompositions that can be used include polymeric substances and waxes.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Alternatively, pharmaceutically acceptable compositions described hereinmay be administered in the form of suppositories for rectal or vaginaladministration. These can be prepared by mixing the compounds of thepresent disclosure with suitable non-irritating excipients or carriersthat are solid at room temperature but liquid at body (e.g. rectal orvaginal) temperature and therefore will melt in the rectum or vaginalcavity to release the active compound. Such materials include cocoabutter, a suppository wax (e.g., beeswax) and polyethylene glycols.

Pharmaceutically acceptable compositions described herein may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Topicalapplication for the lower intestinal tract can be effected in a rectalsuppository formulation (see above) or in a suitable enema formulation.

Dosage forms for topical or transdermal administration of a providedcompound include ointments, pastes, creams, lotions, gels, powders,solutions, sprays, inhalants or patches. The active component is admixedunder sterile conditions with a pharmaceutically acceptable carrier andany needed preservatives or buffers as may be required. Ophthalmicformulations, ear drops, and eye drops are also contemplated as beingwithin the scope of this disclosure. Additionally, the presentdisclosure contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

For topical applications, provided pharmaceutically acceptablecompositions may be formulated in a suitable ointment containing theactive component suspended or dissolved in one or more carriers.Carriers for topical administration of compounds of this disclosureinclude, but are not limited to, mineral oil, liquid petrolatum, whitepetrolatum, propylene glycol, polyoxyethylene, polyoxypropylenecompound, emulsifying wax and water. Alternatively, providedpharmaceutically acceptable compositions can be formulated in a suitablelotion or cream containing the active components suspended or dissolvedin one or more pharmaceutically acceptable carriers. Suitable carriersinclude, but are not limited to, mineral oil, sorbitan monostearate,polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol,benzyl alcohol and water.

For ophthalmic use, provided pharmaceutically acceptable compositionsmay be formulated as micronized suspensions in isotonic, pH adjustedsterile saline, or, preferably, as solutions in isotonic, pH adjustedsterile saline, either with or without a preservative such asbenzylalkonium chloride. Alternatively, for ophthalmic uses, thepharmaceutically acceptable compositions may be formulated in anointment such as petrolatum.

Pharmaceutically acceptable compositions of this disclosure may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

Most preferably, pharmaceutically acceptable compositions of thisdisclosure are formulated for oral administration.

Identification and/or Characterization of Compounds and/or Compositions

Among other things, the present disclosure provides various technologiesfor identification and/or characterization of compounds and/orcompositions as described herein. For example, the present disclosureprovides various assays for assessing SARM1 inhibitory activity, andspecifically for assessing SARM1 inhibitory activity.

In some embodiments, performance of one or more compounds orcompositions of interest in an assay as described herein is comparedwith that of an appropriate reference. For example, in some embodiments,a reference may be absence of the relevant compound or composition.Alternatively or additionally, in some embodiments, a reference may bepresence of an alternative compound or composition, e.g., whichalternative compound or composition has known performance (e.g., as apositive control or a negative control, as is understood in the art) inthe relevant assay. In some embodiments, a reference may be analternative but comparable set of conditions (e.g., temperature, pH,salt concentration, etc.). In some embodiments, a reference may beperformance of the compound or composition with respect to a SARM1variant.

Still further alternatively or additionally, in some embodiments,performance of one or more compounds or compositions of interest in anassay as described herein may be assessed in the presence of anappropriate reference compound or composition, for example, so thatability of the compound or composition to compete with the reference isdetermined.

In some embodiments, a plurality of compounds or compositions ofinterest may be subjected to analysis in a particular assay and/orcompared with the same reference. In some embodiments, such a pluralityof compounds or compositions may be or include a set of compounds orcompositions that is considered to be a “library” because multiplemembers share one or more features (e.g., structural elements, sourceidentity, synthetic similarities, etc.).

Certain exemplary assays that may be useful in the practice of thepresent disclosure are exemplified in the Examples below. Those skilledin the art, reading the present disclosure, will be aware that useful orrelevant systems for identifying and/or characterizing compounds and/orcompositions in accordance with the present disclosure are not limitedto those included in the Examples, or otherwise discussed below.

In some embodiments, compounds and/or compositions may be identifiedbased on and/or characterized by one or more activities orcharacteristics such as, for example: promoting axonal integrity,cytoskeletal stability, and/or neuronal survival. In some embodiments,provided SARM1 inhibitors inhibit catabolism of NAD+ to by SARM1. Insome embodiments, provided SARM1 inhibitors slow the rate of NAD+catabolism.

In some embodiments, provided SARM1 inhibitors reduce or inhibit bindingof NAD+ by SARM1. In some embodiments, provided SARM1 inhibitors bind toSARM1 within a pocket comprising one or more catalytic residues (e.g., acatalytic cleft of SARM1). Examples of such catalytic residues includethe glutamic acid at position 642 (E642).

In some embodiments, provided SARM1 inhibitors disrupt and/or preventmultimerization of the TIR1 domain of SARM1. In some embodiments,provided SARM1 inhibitors disrupt the multimerization of the SAMdomains. In some embodiments, provided SARM1 inhibitors disrupt theaxonal signaling cascade that leads to depletion of NAD+.

In some embodiments, the present disclosure provides assays useful foridentifying and/or characterizing one or more activities and/orcharacteristics of compound and/or compositions of interest. Forexample, in some embodiments, the present disclosure provides in vitro,cellular, and/or in vivo systems for assessing one or more suchactivities and/or characteristics.

SARM1 Activity Assays

In some embodiments, a method of identifying a SARM1 inhibitorcomprises: a) providing a mixture comprising i) a mutant or fragment ofSARM1, ii) NAD+ and iii) a candidate inhibitor, wherein the mutant orfragment has constitutive activity; b) incubating the mixture; c)quantifying NAD+ in the mixture after the incubating; and d) identifyingthe candidate inhibitor compound as an inhibitor if the amount of NAD+is greater than that of a control mixture that does not contain thecandidate inhibitor.

In some embodiments, provided are methods of identifying a SARM1inhibitor, comprising: a) providing a mixture comprising i) afull-length SARM1, ii) NAD+ and iii) a candidate inhibitor, wherein thefull-length SARM1 has constitutive activity; b) incubating the mixture;c) quantifying NAD+ and ADPR (or cADPR) in the mixture after theincubating; d) determining the molar ratio of NAD+:ADPR (or cADPR); ande) identifying the candidate inhibitor compound as an inhibitor if themolar ratio is greater than that of a control mixture that does notcontain the candidate inhibitor.

In some embodiments, provided are methods of identifying a SARM1inhibitor, comprising: a) providing a mixture comprising a solid supportto which is bound i) a full-length SARM1 and at least one tag, ii) NAD+,and iii) a candidate inhibitor; b) incubating the mixture; c)quantifying the NAD+ after the incubating; and d) identifying thecandidate inhibitor compound as a SARM1 inhibitor if the concentrationof NAD+ is greater than that of a control.

SARM1 Binding Assays

In some embodiments, the efficacy of provided SARM1 inhibitors can bedetermined according to, e.g., the assays described in WO 2018/057989,published on Mar. 29, 2018, which is hereby incorporated by reference inits entirety. In some embodiments, the provided SARM1 inhibitors can beapplied to a solution containing SARM1 or a fragment thereof. In someembodiments, the provided SARM1 inhibitors can be applied to an in vitrosystem. In some embodiments, the provided SARM1 inhibitors can beapplied to an in vivo. In some embodiments, the provided SARM1inhibitors can be applied to a patient. In some embodiments, a SARM1inhibitor can be mixed with SARM1 or fragment thereof that has beenlabeled with an epitope tag. In some embodiments, the amount of boundSARM1 inhibitor can be compared to the amount of unbound SARM1inhibitor, yielding the affinity for the SARM1 inhibitor.

In some embodiments, the mutant or fragment of SARM1 is a SAM-TIRfragment having constitutive activity. Fragments of SARM1 havingconstitutive activity include, for example and without limitation, aSARM1 with the autoinhibitory domain deleted; at least one pointmutation of SARM1 that renders the autoinhibitory domain inactive; afragment of SARM1 containing a TIR domain; or a fragment of SARM1consisting of SAM and TIR domains. In some embodiments a SARM1polypeptide can include one or more additional amino acid sequences thatcan act as tags, such as a His tag, a streptavidin tag, or a combinationthereof. In some embodiments a SARM1 polypeptide can include a tag atthe amino terminus, at the carboxy terminus, or a combination thereof.In some embodiments, SARM1 or fragment thereof labeled with an epitopetag can be used to measure the binding efficacy of provided SARM1inhibitors.

Purification of SARM1-TIR Domains

In some embodiments, a SARM1-TIR domain can be engineered with variousprotein, or epitope, tags that can be useful, for example, inpurification. In some embodiments, the present disclosure also providesfor a NRK1-HEK293T cell line comprising HEK293T cells transformed with aNicotinamide Riboside Kinase 1 (NRK1). In some embodiments, HEK293Tcells transformed or transfected with a DNA sequence encodingNicotinamide Riboside Kinase 1 (NRK1). In some embodiments, the DNAencoding NRK1 can be genomic or cDNA. In some embodiments, HEK293T cellsare stably or transiently transfected with DNA encoding NRK1 from asource exogenous to the host cell. In some embodiments, HEK293T cellsare stably or transiently transfected with DNA encoding NRK1 such thatthe cells express NRK1 at an elevated level compared to control cells.In some embodiments, DNA encoding NRK1 is under the control of one ormore exogenous regulatory DNA sequences such as a promoter, an enhanceror a combination thereof. In some embodiments, a combination of a DNAsequences encoding NRK1 and regulatory sequences is a non-naturallyoccurring combination. In some embodiments, DNA encoding NRK1, eithergenomic or cDNA, comprises an expression vector such as an FCIVexpression vector. In some embodiments, DNA encoding NRK1 is derivedfrom genomic DNA or cDNA from a vertebrate or invertebrate species suchas, but not limited to, human, mouse, zebrafish or a Drosophila. In someconfigurations, the NRK1 DNA is a human NRK1 DNA.

Applications and Uses

The present disclosure provides a variety of uses and applications forcompounds and/or compositions as described herein, for example in lightof their activities and/or characteristics as described herein. In someembodiments, such uses may include therapeutic and/or diagnostic uses.Alternatively, in some embodiments such uses may include research,production, and/or other technological uses.

In one aspect, the present disclosure provides methods comprisingadministering one or more compounds of Formula I or Formula II to asubject, e.g., to treat, prevent, or reduce the risk of developing oneor more conditions characterized by axonal degeneration. In some suchembodiments, the compound of Formula I or Formula II is a SARM1inhibitor.

Another embodiment of the present disclosure relates to a method ofinhibiting SARM1 activity in a patient comprising steps of administeringto said patient a provided compound, or a composition comprising saidcompound.

Inhibition of enzymes in a biological sample is useful for a variety ofpurposes that are known to one of skill in the art. Examples of suchpurposes include, but are not limited to biological assays, geneexpression studies, and biological target identification.

In certain embodiments, the present disclosure relates to a method oftreating axonal degeneration in a biological sample comprising the stepof contacting said biological sample with a compound or composition ofFormula I or Formula II. In some embodiments, one or more compoundsand/or compositions as described herein are useful, for example as amethod of for inhibiting the degradation of neurons derived from asubject. In some embodiments, one or more compounds and/or compositionsas described herein, are useful for inhibiting the degeneration of aneuron, or portion thereof, cultured in vitro. In some embodiments, oneor more compounds and/or compositions as described herein, are useful asstabilizing agents to promote in vitro neuronal survival.

In some embodiments, provided compounds and/or compositions inhibitNADase activity of SARM1. Alternatively or additionally, in someembodiments, provided compounds alleviate one or more attributes ofneurodegeneration. In some embodiments, the present disclosure providesmethods of treating a neurodegenerative disease or disorder associatedwith axonal degeneration.

In some embodiments, one or more compounds and/or compositions asdescribed herein are useful, for example, in the practice of medicine.In some embodiments, one or more compounds and/or compositions asdescribed herein are useful, for example, to treat, prevent, orameliorate axonal degeneration (e.g., one or more features orcharacteristics thereof). In some embodiments, one or more compoundsand/or compositions as described herein are useful, for example toinhibit axonal degeneration, including axonal degeneration that resultsfrom reduction or depletion of NAD+. In some embodiments, one or morecompounds and/or compositions as described herein are useful, forexample to prevent the axon distal to an axonal injury fromdegenerating.

In some embodiments, one or more compounds and/or compositions asdescribed herein are useful, for example as a method for inhibiting thedegradation of a peripheral nervous system neuron or a portion thereof.In some embodiments, one or more compounds and/or compositions asdescribed herein are useful, for example as a method for inhibiting orpreventing degeneration of a central nervous system (neuron) or aportion thereof. In some embodiments, one or more compounds orcompositions as described herein is characterized that, whenadministered to a population of subjects, reduces one or more symptomsor features of neurodegeneration. For example, in some embodiments, arelevant symptom or feature may be selected from the group consisting ofextent, rate, and/or timing of neuronal disruption.

In certain embodiments, the present disclosure provides compounds thatare useful, for example, as analytical tools, as probes in biologicalassays, or as therapeutic agents in accordance with the presentdisclosure. Compounds provided by this disclosure are also useful forthe study of SARM1 activity in biological and pathological phenomena andthe comparative evaluation of new SARM1 activity inhibitors in vitro orin vivo. In certain embodiments, the present disclosure provides assaysfor identifying and/or characterizing compounds and/or compositionsprovided herein. In some embodiments, provided assays utilize particularreagents and/or systems (e.g., certain vector constructs and/orpolypeptides) useful in assaying SARM1 activity. For example, in someembodiments, provided assays may utilize, for example, a SAM-TIR inwhich the SARM1 N-terminal auto-inhibitory domain is deleted, and/or oneor more tagged versions of a TIR domain.

In some embodiments, one or more compounds and/or compositions asdescribed herein are useful, for example as a method of for inhibitingthe degradation of neurons derived from a subject. In some embodiments,one or more compounds and/or compositions as described herein, areuseful for inhibiting the degeneration of a neuron, or portion thereof,cultured in vitro. In some embodiments, one or more compounds and/orcompositions as described herein, are useful as stabilizing agents topromote in vitro neuronal survival.

In some embodiments, one or more compounds and/or compositions asdescribed herein are useful, for example in affecting biomarkersassociated with neurodegeneration. In some embodiments, changes inbiomarkers can be detected systemically or with a sample of cerebralspinal fluid (CSF), plasma, serum, and/or tissue from a subject. In someembodiments, one or more compounds and/or compositions can be used toaffect a change in the concentration of neurofilament protein light(NF-L) and/or neurofilament protein heavy (NF-H) contained the cerebralspinal fluid of a subject. In some embodiments, one or more compoundsand/or compositions as described herein can affect constitutive NADand/or cADPR levels in neurons and/or axons.

In some embodiments, one or more compounds and/or compositions asdescribed herein can affect a detectable change in the levels of one ormore neurodegeneration-associated proteins in a subject. Such proteinsinclude, but are not limited to, albumin, amyloid-β (Aβ)38, Aβ40, Aβ42,glial fibrillary acid protein (GFAP), heart-type fatty acid bindingprotein (hFABP), monocyte chemoattractin protein (MCP)-1, neurogranin,neuron specific enolayse (NSE), soluble amyloid precursor protein(sAPP)α, sAPPβ, soluble triggering receptor expressed on myeloid cells(sTREM) 2, phospho-tau, and/or total-tau. In some embodiments, one ormore compounds and/or compositions as described herein can affect achange in cytokines and/or chemokines, including, but not limited to,Ccl2, Ccl7, Ccl12, Csf1, and/or Il6.

Diseases, Disorders, and Conditions

In some embodiments, compounds and/or compositions as described hereinmay be administered to subjects suffering from one or more diseases,disorders, or conditions.

In some embodiments, the condition is an acute condition. In someembodiments, the condition is a chronic condition.

In some embodiments, the condition is characterized by axonaldegeneration in the central nervous system, the peripheral nervoussystem, the optic nerve, the cranial nerves, or a combination thereof.

In some embodiments, the condition is or comprises acute injury to thecentral nervous system, e.g., injury to the spinal cord and/or traumaticbrain injury. In some embodiments, the condition is or comprises achronic injury to the central nervous system, e.g., injury to the spinalcord, traumatic brain injury, and/or traumatic axonal injury. In someembodiments, the condition is or comprises chronic traumaticencephalopathy (CTE).

In some embodiments, the condition is a chronic condition affecting thecentral nervous system, e.g., Parkinson's disease, amyotrophic lateralsclerosis, multiple sclerosis, or Huntington disease, Alzheimer'sdisease.

In some embodiments, the condition is an acute peripheral neuropathy.Chemotherapy-induced peripheral neuropathy (CIPN) is an example of anacute peripheral neuropathy. CIPN can be associated with various drugs,such as, but not limited to, thalidomide, epothilones (e.g.,ixabepilone), taxanes (e.g., paclitaxel and docetaxel), vinca alkaloids(e.g., vinblastine, vinorelbine, vincristine, and vindesine), proteasomeinhibitors (e.g., bortezomib), platinum-based drugs (e.g., cisplatin,oxaliplatin, and carboplatin).

In some embodiments, the condition is a chronic condition affecting theperipheral nervous system, e.g., diabetic neuropathy, HIV neuropathy,Charcot Marie Tooth disease, or amyotrophic lateral sclerosis.

In some embodiments, the condition is an acute condition affecting theoptic nerve, e.g., acute optic neuropathy (AON) or acute angle closureglaucoma.

In some embodiments, the condition is a chronic condition affecting theoptic nerve, e.g., Leber's congenital amaurosis, Leber's hereditaryoptic neuropathy, primary open angle glaucoma, and autosomal dominantoptic atrophy.

In some embodiments, one or more compounds and/or compositions asdescribed herein are useful, for example, to treat one or moreneurodegenerative diseases, disorders or conditions selected from thegroup consisting of neuropathies or axonopathies. In some embodiments,one or more compounds and/or compositions as described herein areuseful, for example to treat a neuropathy or axonopathy associated withaxonal degeneration. In some embodiments, a neuropathy associated withaxonal degeneration is a hereditary or congenital neuropathy oraxonopathy. In some embodiments, a neuropathy associated with axonaldegeneration results from a de novo or somatic mutation. In someembodiments, a neuropathy associated with axonal degeneration isselected from a list contained herein. In some embodiments, a neuropathyor axonopathy is associated with axonal degeneration, including, but notlimited to Parkinson's disease, non-Parkinson's disease, Alzheimer'sdisease, Herpes infection, diabetes, amyotrophic lateral sclerosis, ademyelinating disease, ischemia or stroke, chemical injury, thermalinjury, and AIDS.

In some embodiments, one or more compounds or compositions as describedherein is characterized that, when administered to a population ofsubjects, reduces one or more symptoms or features of neurodegeneration.For example, in some embodiments, a relevant symptom or feature may beselected from the group consisting of extent, rate, and/or timing ofneuronal disruption. In some embodiments, neuronal disruption may be orcomprise axonal degradation, loss of synapses, loss of dendrites, lossof synaptic density, loss of dendritic arborization, loss of axonalbranching, loss of neuronal density, loss of myelination, loss ofneuronal cell bodies, loss of synaptic potentiation, loss ofaction-potential potentiation, loss of cytoskeletal stability, loss ofaxonal transport, loss of ion channel synthesis and turnover, loss ofneurotransmitter synthesis, loss of neurotransmitter release andreuptake capabilities, loss of axon-potential propagation, neuronalhyperexitability, and/or neuronal hypoexcitability. In some embodiments,neuronal disruption is characterized by an inability to maintain anappropriate resting neuronal membrane potential. In some embodiments,neuronal disruption is characterized by the appearance of inclusionbodies, plaques, and/or neurofibrillary tangles. In some embodiments,neuronal disruption is characterized by the appearance of stressgranules. In some embodiments, neuronal disruption is characterized bythe intracellular activation of one or more members of thecysteine-aspartic protease (Caspase) family. In some embodiments,neuronal disruption is characterized by a neuron undergoing programedcell death (e.g. apoptosis, pyroptosis, ferroapoptosis, and/or necrosis)and/or inflammation.

In some embodiments, the neurodegenerative or neurological disease ordisorder is associated with axonal degeneration, axonal damage,axonopathy, a demyelinating disease, a central pontine myelinolysis, anerve injury disease or disorder, a metabolic disease, a mitochondrialdisease, metabolic axonal degeneration, axonal damage resulting from aleukoencephalopathy or a leukodystrophy. In some embodiments, theneurodegenerative or neurological disease or disorder is selected fromthe group consisting of spinal cord injury, stroke, multiple sclerosis,progressive multifocal leukoencephalopathy, congenital hypomyelination,encephalomyelitis, acute disseminated encephalomyelitis, central pontinemyelolysis, osmotic hyponatremia, hypoxic demyelination, ischemicdemyelination, adrenoleukodystrophy, Alexander's disease, Niemann-Pickdisease, Pelizaeus Merzbacher disease, periventricular leukomalacia,globoid cell leukodystrophy (Krabbe's disease), Wallerian degeneration,optic neuritis, transverse myelitis, amyotrophic lateral sclerosis (ALS,Lou Gehrig's disease), Huntington's disease, Alzheimer's disease,Parkinson's disease, Tay-Sacks disease, Gaucher's disease, HurlerSyndrome, traumatic brain injury, post radiation injury, neurologiccomplications of chemotherapy (chemotherapy induced neuropathy; CIPN),neuropathy, acute ischemic optic neuropathy, vitamin B12 deficiency,isolated vitamin E deficiency syndrome, Bassen-Kornzweig syndrome,Glaucoma, Leber's hereditary optic atrophy (neuropathy), Lebercongenital amaurosis, neuromyelitis optica, metachromaticleukodystrophy, acute hemorrhagic leukoencephalitis, trigeminalneuralgia, Bell's palsy, cerebral ischemia, multiple system atrophy,traumatic glaucoma, tropical spastic paraparesis human T-lymphotropicvirus 1 (HTLV-1) associated myelopathy, west Nile virus encephalopathy,La Crosse virus encephalitis, Bunyavirus encephalitis, pediatric viralencephalitis, essential tremor, Charcot-Marie-Tooth disease, motorneuron disease, spinal muscular atrophy (SMA), hereditary sensory andautonomic neuropathy (HSAN), adrenomyeloneuropathy, progressive supranuclear palsy (PSP), Friedrich's ataxia, hereditary ataxias, noiseinduced hearing loss, congenital hearing loss, Lewy Body Dementia,frontotemporal dementia, amyloidosis, diabetic neuropathy, HIVneuropathy, enteric neuropathies and axonopathies, Guillain-Barresyndrome, severe acute motor axonal neuropathy (AMAN), Creutzfeldt-Jakobdisease, transmissible spongiform encephalopathy, spinocerebellarataxias, pre-eclampsia, hereditary spastic paraplegias, spasticparaparesis, familial spastic paraplegia, French settlement disease,Strumpell-Lorrain disease, and non-alcoholic steatohepatitis (NASH).

In some embodiments, the present disclosure provides inhibitors of SARM1activity for treatment of neurodegenerative or neurological diseases ordisorders that involve axon degeneration or axonopathy. The presentdisclosure also provides methods of using inhibitors of SARM1 activityto treat, prevent or ameliorate axonal degeneration, axonopathies andneurodegenerative or neurological diseases or disorders that involveaxonal degeneration.

In some embodiments, the present disclosure provides methods of treatingneurodegenerative or neurological diseases or disorders related toaxonal degeneration, axonal damage, axonopathies, demyelinatingdiseases, central pontine myelinolysis, nerve injury diseases ordisorders, metabolic diseases, mitochondrial diseases, metabolic axonaldegeneration, axonal damage resulting from a leukoencephalopathy or aleukodystrophy.

In some embodiments, neuropathies and axonopathies include any diseaseor condition involving neurons and/or supporting cells, such as forexample, glia, muscle cells or fibroblasts, and, in particular, thosediseases or conditions involving axonal damage. Axonal damage can becaused by traumatic injury or by non-mechanical injury due to diseases,conditions, or exposure to toxic molecules or drugs. The result of suchdamage can be degeneration or dysfunction of the axon and loss offunctional neuronal activity. Disease and conditions producing orassociated with such axonal damage are among a large number ofneuropathic diseases and conditions. Such neuropathies can includeperipheral neuropathies, central neuropathies, and combinations thereof.Furthermore, peripheral neuropathic manifestations can be produced bydiseases focused primarily in the central nervous systems and centralnervous system manifestations can be produced by essentially peripheralor systemic diseases.

In some embodiments, a peripheral neuropathy can involve damage to theperipheral nerves, and/or can be caused by diseases of the nerves or asthe result of systemic illnesses. Some such diseases can includediabetes, uremia, infectious diseases such as AIDs or leprosy,nutritional deficiencies, vascular or collagen disorders such asatherosclerosis, and autoimmune diseases such as systemic lupuserythematosus, scleroderma, sarcoidosis, rheumatoid arthritis, andpolyarteritis nodosa. In some embodiments, peripheral nerve degenerationresults from traumatic (mechanical) damage to nerves as well as chemicalor thermal damage to nerves. Such conditions that injure peripheralnerves include compression or entrapment injuries such as glaucoma,carpal tunnel syndrome, direct trauma, penetrating injuries, contusions,fracture or dislocated bones; pressure involving superficial nerves(ulna, radial, or peroneal) which can result from prolonged use ofcrutches or staying in one position for too long, or from a tumor;intraneural hemorrhage; ischemia; exposure to cold or radiation orcertain medicines or toxic substances such as herbicides or pesticides.In particular, the nerve damage can result from chemical injury due to acytotoxic anticancer agent such as, for example, taxol, cisplatinin, aproteasome inhibitor, or a vinca alkaloid such as vincristine. Typicalsymptoms of such peripheral neuropathies include weakness, numbness,paresthesia (abnormal sensations such as burning, tickling, pricking ortingling) and pain in the arms, hands, legs and/or feet. In someembodiments, a neuropathy is associated with mitochondrial dysfunction.Such neuropathies can exhibit decreased energy levels, i.e., decreasedlevels of NAD and ATP.

In some embodiments, peripheral neuropathy is a metabolic and endocrineneuropathy which includes a wide spectrum of peripheral nerve disordersassociated with systemic diseases of metabolic origin. These diseasesinclude, for example, diabetes mellitus, hypoglycemia, uremia,hypothyroidism, hepatic failure, polycythemia, amyloidosis, acromegaly,porphyria, disorders of lipid/glycolipid metabolism, nutritional/vitamindeficiencies, and mitochondrial disorders, among others. The commonhallmark of these diseases is involvement of peripheral nerves byalteration of the structure or function of myelin and axons due tometabolic pathway dysregulation.

In some embodiments, neuropathies include optic neuropathies such asglaucoma; retinal ganglion degeneration such as those associated withretinitis pigmentosa and outer retinal neuropathies; optic nerveneuritis and/or degeneration including that associated with multiplesclerosis; traumatic injury to the optic nerve which can include, forexample, injury during tumor removal; hereditary optic neuropathies suchas Kjer's disease and Leber's hereditary optic neuropathy; ischemicoptic neuropathies, such as those secondary to giant cell arteritis;metabolic optic neuropathies such as neurodegenerative diseasesincluding Leber's neuropathy mentioned earlier, nutritional deficienciessuch as deficiencies in vitamins B12 or folic acid, and toxicities suchas due to ethambutol or cyanide; neuropathies caused by adverse drugreactions and neuropathies caused by vitamin deficiency. Ischemic opticneuropathies also include non-arteritic anterior ischemic opticneuropathy.

In some embodiments neurodegenerative diseases that are associated withneuropathy or axonopathy in the central nervous system include a varietyof diseases. Such diseases include those involving progressive dementiasuch as, for example, Alzheimer's disease, senile dementia, Pick'sdisease, and Huntington's disease; central nervous system diseasesaffecting muscle function such as, for example, Parkinson's disease,motor neuron diseases and progressive ataxias such as amyotrophiclateral sclerosis; demyelinating diseases such as, for example multiplesclerosis; viral encephalitides such as, for example, those caused byenteroviruses, arboviruses, and herpes simplex virus; and priondiseases. Mechanical injuries such as glaucoma or traumatic injuries tothe head and spine can also cause nerve injury and degeneration in thebrain and spinal cord. In addition, ischemia and stroke as well asconditions such as nutritional deficiency and chemical toxicity such aswith chemotherapeutic agents can cause central nervous systemneuropathies.

In some embodiments, the present disclosure provides a method oftreating a neuropathy or axonopathy associated with axonal degeneration.In some such embodiments, a neuropathy or axonopathy associated withaxonal degeneration can be any of a number of neuropathies oraxonopathies such as, for example, those that are hereditary orcongenital or associated with Parkinson's disease, Alzheimer's disease,Herpes infection, diabetes, amyotrophic lateral sclerosis, ademyelinating disease, ischemia or stroke, chemical injury, thermalinjury, and AIDS. In addition, neurodegenerative diseases not mentionedabove as well as a subset of the above mentioned diseases can also betreated with the methods of the present disclosure. Such subsets ofdiseases can include Parkinson's disease or non-Parkinson's diseases, orAlzheimer's disease.

Subjects

In some embodiments, compounds and/or compositions as described hereinare administered to subjects suffering from or susceptible to a disease,disorder or condition as described herein; in some embodiments, such adisease, disorder or condition is characterized by axonal degeneration,such as one of the conditions mentioned herein.

In some embodiments, a subject to whom a compound or composition isadministered as described herein exhibits one or more signs or symptomsassociated with axonal degeneration; in some embodiments, the subjectdoes not exhibit any signs or symptoms of neurodegeneration.

In some embodiments, provided methods comprise administering a compoundof Formula I or Formula II to a patient in need thereof. In some suchembodiments, the patient is at risk of developing a conditioncharacterized by axonal degeneration. In some embodiments, the patienthas a condition characterized by axonal degeneration. In someembodiments, the patient has been diagnosed with a conditioncharacterized by axonal degeneration.

In some embodiments, provided methods comprise administering acomposition as described herein to a patient population of in needthereof. In some embodiments, the population is drawn from individualswho engage in activities where the potential for traumatic neuronalinjury is high. In some embodiments, the population is drawn fromathletes who engage in contact sports or other high-risk activities.

In some embodiments, the subject is at risk of developing a conditioncharacterized by axonal degeneration. In some embodiments, the subjectis identified as being at risk of axonal degeneration, e.g., based onthe subject's genotype, a diagnosis of a condition associated withaxonal degeneration, and/or exposure to an agent and/or a condition thatinduces axonal degeneration.

In some embodiments, the patient is at risk of developing aneurodegenerative disorder. In some embodiments, the patient is elderly.In some embodiments, the patient is known to have a genetic risk factorfor neurodegeneration. In some embodiments, the patient has a familyhistory of neurodegenerative disease. In some embodiments, the patientexpresses one or more copies of a known genetic risk factor forneurodegeneration. In some embodiments, the patient is drawn from apopulation with a high incidence of neurodegeneration. In someembodiments, the patient has a hexanucleotide repeat expansion inchromosome 9 open reading frame 72. In some embodiments, the patient hasone or more copies of the ApoE4 allele.

In some embodiments, subjects to which a compound or composition asdescribed herein is administered may be or comprise subjects sufferingfrom or susceptible to a neurodegenerative disease, disorder orcondition. In some embodiments, a neurodegenerative disease, disorder orcondition may be or comprise a traumatic neuronal injury. In someembodiments, a traumatic neuronal injury is blunt force trauma, aclosed-head injury, an open head injury, exposure to a concussive and/orexplosive force, a penetrating injury in to the brain cavity orinnervated region of the body. In some embodiments, a traumatic neuronalinjury is a force which causes the axons to deform, stretch, crush orsheer.

In some embodiments, the subject engages in an activity identified as arisk factor for neuronal degradation, e.g., a subject that engages incontact sports or occupations with a high chance for traumatic neuronalinjury.

For example, the subject may be a patient who is receiving, or isprescribed, a chemotherapy associated with peripheral neuropathy.Examples of chemotherapeutic agents include, but not limited to,thalidomide, epothilones (e.g., ixabepilone), taxanes (e.g., paclitaxeland docetaxel), vinca alkaloids (e.g., vinblastine, vinorelbine,vincristine, and vindesine), proteasome inhibitors (e.g., bortezomib),platinum-based drugs (e.g., cisplatin, oxaliplatin, and carboplatin).

In some embodiments, provided methods comprise administering acomposition as described herein to a patient or patient population basedon the presence or absence of one or more biomarkers. In someembodiments, provided methods further comprise monitoring the level of abiomarker in a patient or patient population and adjusting the dosingregimen accordingly.

Dosing

Those of skill in the art will appreciate that, in some embodiments, theexact amount of a particular compound included in and/or delivered byadministration of a pharmaceutical composition or regimen as describedherein may be selected by a medical practitioner and may be differentfor different subjects, for example, upon consideration of one or moreof species, age, and general condition of the subject, and/or identityof the particular compound or composition, its mode of administration,and the like. Alternatively, in some embodiments, the amount of aparticular compound included in and/or delivered by administration of apharmaceutical composition or regimen as described herein may bestandardized across a relevant patient population (e.g., all patients,all patients of a particular age or stage of disease or expressing aparticular biomarker, etc.).

A provided compound or composition of the present disclosure ispreferably formulated in dosage unit form for ease of administration anduniformity of dosage. The expression “dosage unit form” as used hereinrefers to a physically discrete unit of agent appropriate for thepatient to be treated. It will be understood, however, that the totaldaily usage of a provided compound or composition of the presentdisclosure will be decided by the attending physician within the scopeof sound medical judgment. The specific effective dose level for anyparticular patient or organism will depend upon a variety of factorsincluding the disorder being treated and the severity of the disorder;the clinical condition of the individual patient; the cause of thedisorder; the activity of the specific compound employed; the specificcomposition employed; the age, body weight, general health, sex and dietof the patient; the time of administration, delivery site of the agent,route of administration, and rate of excretion of the specific compoundemployed; the duration of the treatment; drugs used in combination orcoincidental with the specific compound employed, and like factors wellknown in the medical arts. The effective amount of the compound to beadministered will be governed by such considerations, and is the minimumamount necessary to inhibit SARM1 activity as required to prevent ortreat the undesired disease or disorder, such as for example,neurodegeneration or traumatic neural injury.

A pharmaceutically acceptable composition of this disclosure can beadministered to humans and other animals orally, rectally,intravenously, parenterally, intracisternally, intravaginally,intraperitoneally, topically (as by powders, ointments, or drops),bucally, as an oral or nasal spray, or the like, depending on theseverity of the disease, disorder or infection being treated. The dailydose is, in certain embodiments, given as a single daily dose or individed doses two to six times a day, or in sustained release form. Thisdosage regimen may be adjusted to provide the optimal therapeuticresponse. The compounds may be administered on a regimen of 1 to 4 timesper day, preferably once or twice per day.

In some embodiments, compositions of the present disclosure may beadministered orally, parenterally, by inhalation spray, topically,rectally, nasally, buccally, vaginally or via an implanted reservoir.The term “parenteral” as used herein includes subcutaneous, intravenous,intramuscular, intra-articular, intra-synovial, intrasternal,intrathecal, intrahepatic, intradermal, intraocular, intralesional andintracranial injection or infusion techniques. Preferably, thecompositions are administered orally, intraperitoneally orintravenously.

In some embodiments, pharmaceutically acceptable compositions of thisdisclosure may also be administered topically, especially when thetarget of treatment includes areas or organs readily accessible bytopical application, including diseases of the eye, the skin, or thelower intestinal tract. Suitable topical formulations are readilyprepared for each of these areas or organs.

Most preferably, pharmaceutically acceptable compositions of thisdisclosure are formulated for oral administration. Such formulations maybe administered with or without food. In some embodiments,pharmaceutically acceptable compositions of this disclosure areadministered without food. In other embodiments, pharmaceuticallyacceptable compositions of this disclosure are administered with food.

Those additional agents may be administered separately from a providedcompound or composition thereof, as part of a multiple dosage regimen.Alternatively, those agents may be part of a single dosage form, mixedtogether with a provided compound in a single composition. Ifadministered as part of a multiple dosage regime, the two active agentsmay be submitted simultaneously, sequentially or within a period of timefrom one another, normally within five hours from one another.

It should also be understood that a specific dosage and treatmentregimen for any particular patient may depend upon a variety of factors,including the activity of the specific compound employed, the age, bodyweight, general health, sex, diet, time of administration, rate ofexcretion, drug combination, and the judgment of the treating physicianand the severity of the particular disease being treated. In someembodiments, the amount of a compound of the present disclosure in thecomposition will also depend upon the particular compound in thecomposition.

In some embodiments, SARM1 inhibition as described herein may beutilized in combination with one or more other therapies to treat arelevant disease, disorder, or condition. In some embodiments, dosing ofa SARM1 inhibitor is altered when utilized in combination therapy ascompared with when administered as monotherapy; alternatively oradditionally, in some embodiments, a therapy that is administered incombination with SARM1 inhibition as described herein is administeredaccording to a regimen or protocol that differs from its regimen orprotocol when administered alone or in combination with one or moretherapies other than SARM1 inhibition. In some embodiments, compositionswhich comprise an additional therapeutic agent, that additionaltherapeutic agent and a provided compound may act synergistically. Insome embodiments, one or both therapies utilized in a combinationregimen is administered at a lower level or less frequently than when itis utilized as monotherapy.

In some embodiments, compounds and/or compositions described herein areadministered with a chemotherapeutic agent including, but not limitedto, alkylating agents, anthracyclines, taxanes, epothilones, histonedeacetylase inhibitors, topoisomerase inhibitors, kinase inhibitors,nucleotide analogs, peptide antibiotics, platinum-based agents,retinoids, vinca alkaloids and derivatives. In some embodiments,compounds and/or compositions described herein are administered incombination with PARP inhibitors.

EXEMPLIFICATION

The present teachings including descriptions provided in the Examplesthat are not intended to limit the scope of any claim. Unlessspecifically presented in the past tense, inclusion in the Examples isnot intended to imply that the experiments were actually performed. Thefollowing non-limiting examples are provided to further illustrate thepresent teachings. Those of skill in the art, in light of the presentdisclosure, will appreciate that many changes can be made in thespecific embodiments that are disclosed and still obtain a like orsimilar result without departing from the spirit and scope of thepresent teachings.

Example 1. Preparation of the Compounds According to the Invention

General Synthetic Methods

The compounds according to the present invention and their intermediatesmay be obtained using methods of synthesis which are known to the oneskilled in the art and described in the literature of organic synthesis.Preferably, the compounds are obtained in analogous fashion to themethods of preparation explained more fully hereinafter, in particularas described in the experimental section. In some cases, the order incarrying out the reaction steps may be varied. Variants of the reactionmethods that are known to the one skilled in the art but not describedin detail here may also be used.

The general processes for preparing the compounds according to theinvention will become apparent to the one skilled in the art studyingthe following schemes. Starting materials may be prepared by methodsthat are described in the literature, herein, or may be prepared in ananalogous or similar manner. Any functional groups in the startingmaterials or intermediates may be protected using conventionalprotecting groups. These protecting groups may be cleaved again at asuitable stage within the reaction sequence using methods familiar tothe one skilled in the art.

Optimum reaction conditions and reaction times may vary depending on theparticular reactants used. Unless otherwise specified, solvents,temperatures, pressures and other reaction conditions may be readilyselected by one of ordinary skill in the art. Specific procedures areprovided in the Synthetic Examples section. Intermediates and productsmay be purified by chromatography on silica gel, recrystallizationand/or reverse phase HPLC (RHPLC). Discrete enantiomers may be obtainedby resolution of racemic products using chiral HPLC. RHdPLC purificationmethods used anywhere from 0-100% acetonitrile in water containing 0.1%formic acid, 0.1-0.01% TFA, 10 mM aqueous ammonium bicarbonate or 0.2%aqueous ammonium hydroxide and used one of the following columns:

-   -   a) Waters Xbridge C18 10 μm 30×100 mm column    -   b) Waters Sunfire C18 10 μm 30×100 mm column    -   c) Waters Xbridge C18 3.5 μm 50×4.6 mm column    -   d) HALO C18 2.7 μm 30×4.6 mm column    -   e) Waters Sunfire C18 3.5 μm 50×4.6 mm column

LCMS Methods

Analytical LC/MS Analysis Method A:

ESI+/−ion mode 100-1000 Da

Column: XBridge SB-C18, 3.5 μm 4.6×50 mm column

Temperature: 40° C.

Gradient:

10 mM aqueous ammonium Time (min) bicarbonate Acetonitrile Flow (mL/min)0.00 95%  5% 2.0 1.40  5% 95% 2.0 3.00  5% 95% 2.0

Analytical LC/MS Analysis Method B:

ESI+/−ion mode 100-1000 Da

Column: Sunfire C18, 3.5 μm 4.6×50 mm column

Temperature: 50° C.

Gradient:

10 mM aqueous ammonium Time (min) bicarbonate Acetonitrile Flow (mL/min)0.00 95%  5% 2.0 1.40  5% 95% 2.0 3.00  5% 95% 2.0

Analytical LC/MS Analysis Method C:

ESI+/−ion mode 100-1000 Da

Column: XBridge SB-C18, 3.5 μm 4.6×50 mm column

Temperature: 40° C.

Gradient:

10 mM aqueous ammonium Time (min) bicarbonate Acetonitrile Flow (mL/min)0.00 95%  5% 2.0 1.20  5% 95% 2.0

Analytical LC/MS Analysis Method D:

ESI+/−ion mode 100-1000 Da

Column: XBridge SB-C18, 3.5 μm 4.6×50 mm column

Temperature: 40° C.

Gradient:

Time (min) 0.01% TFA in water Acetonitrile Flow (mL/min) 0.00 95%  5%1.8 1.80  5% 95% 1.8 3.00  5% 95% 1.8

Analytical LC/MS Analysis Method E:

ESI+/−ion mode 100-1000 Da

Column: XBridge C18, 3.5 μm 4.6×50 mm column

Temperature: 40° C.

Gradient:

Time (min) 0.01% TFA in water Acetonitrile Flow (mL/min) 0.00 95%  5%2.0 1.30  5% 95% 2.0

Analytical LC/MS Analysis Method F:

ESI+/−ion mode 100-1000 Da

Column: XBridge C18, 3.5 μm 4.6×50 mm column

Temperature: 50° C.

Gradient:

10 mM aqueous ammonium Time (min) bicarbonate Acetonitrile Flow (mL/min)0.00 90% 10% 1.5 1.50  5% 95% 1.5

Analytical LC/MS Analysis Method G:

ESI+/−ion mode 100-1000 Da

Column: XBridge C18, 3.5 μm 4.6×50 mm column

Temperature: 45° C.

Gradient:

Time (min) 0.1% TFA in water Acetonitrile Flow (mL/min) 0.00 95%  5% 1.81.30  5% 95% 1.8 3.00  5% 95% 1.8

Analytical LC/MS Analysis Method H:

ESI+/−ion mode 100-1000 Da

Column: XBridge C18, 3.5 μm 4.6×50 mm column

Temperature: 50° C.

Gradient:

10 mM aqueous ammonium Time (min) bicarbonate Acetonitrile Flow (mL/min)0.00 95%  5% 1.8 1.30  5% 95% 1.8 3.00  5% 95% 1.8

Analytical LC/MS Analysis Method L:

ESI+/−ion mode 100-10001 Da

Column: XBridge C18, 3.5 μm 4.6×50 mm column

Temperature: 45° C.

Gradient:

10 mM aqueous ammonium Time (min) bicarbonate Acetonitrile Flow (mL/min)0.00 90% 10% 1.8 1.40  5% 95% 1.8 3.00  5% 95% 1.8

Analytical LC/MS Analysis Method J:

ESI+/−ion mode 100-1000 Da

Column: Phenomenex Kinetix-XB C18, 1.7 μm 2.1×100 mm column

Temperature: 40° C.

Gradient:

Time Water (0.1% Acetonitrile (0.1% Flow (min) formic acid) formic acid)(mL/min) 0.00 95% 5% 0.6 5.30 0 100 0.6 5.82 95% 5% 0.6 7.00 95% 5% 0.6

SYNTHETIC EXAMPLES Synthetic Example A: Synthesis of Example 76

To a solution of R-1 (80 mg, 0.36 mmol) in DMF (3 mL) is added R-2 (53mg, 0.43 mmol) and K₂CO₃ (149 mg, 1.1 mmol). The mixture is stirred at80° C. for 2 h then cooled, treated with H₂O (10 mL) and extracted withEtOAc (10 mL×3). The organic layer is washed with brine (30 mL×2), driedover Na₂SO₄, and concentrated under reduced pressure. The crude residueis purified by Prep-HPLC to afford Example 76 (72 mg).

The following examples are prepared in similar fashion from theappropriate reagents: Examples 1-6, 8-14, 16-19, 26-27, 35, 47-64,67-69, 75-77, and 82-83.

Synthetic Example B: Synthesis of Example 20

Step 1

A mixture of R-3 (6.0 g, 26.55 mmol) and R-4 (8.2 g, 79.65 mmol) inacetic acid (25 mL) is stirred at 130° C. for 16 h. The reaction mixtureis cooled and concentrated under reduced pressure to remove acetic acid.The residue is diluted with water (200 mL), neutralized with saturatedaqueous NaHCO₃ to pH=7-8 and filtered. The filtrate is acidified withconc. aqueous HCl to pH=1-2, filtered and the solid is dried to affordI-1 (5.4 g, 76%).

Step 2

A mixture of I-1 (2.7 g, 10.0 mmol) and PCl₅ (5.2 g, 25.0 mmol) in POCl₃(20 mL) is stirred at 110° C. for 1.5 h. The reaction mixture is cooledand concentrated under reduced pressure to afford a residue that isdissolved in THE (30 mL). The mixture is added to a solution of ammoniumhydroxide (20 g, 30% aqueous, 171.3 mmol) in THE (10 mL) at 0° C. Afterstirring at 0° C. for 0.5 h, the reaction is concentrated under reducedpressure, filtered, and the resulting solid is washed with water (10 mL)and dried to give a solid.

The solid is dissolved in CH₂Cl₂ (40 mL), cooled to 0° C. then treatedwith TEA (2.14 mL, 15.4 mmol). After 5 min, TFAA (1.3 mL, 9.24 mmol) isadded slowly and the mixture is stirred at 0° C. for 1 h. The mixture isneutralized with NaHCO₃ to pH=7, extracted with CH₂Cl₂ (3×60 mL), driedover Na₂SO₄, filtered and concentrated under reduced pressure. Theresidue is washed with EtOAc (10 mL) and the solid filtered and dried togive I-2 (1.78 g, 68%).

Step 3

A mixture of I-2 (0.4 g, 1.5 mmol), R-5 (0.36 g, 1.8 mmol) and K₂CO₃(1.0 g, 7.5 mmol) in DMF (8 mL) is stirred at 80° C. for 1 h. Thereaction is cooled to room temperature and diluted with EtOAc (150 mL),washed with water (20 mL), brine (4×20 mL), dried over Na₂SO₄, filteredand concentrated under reduced pressure to afford the Boc amineintermediate that is dissolved in CH₂Cl₂ (20 mL). The mixture is treatedwith TFA (5 mL) and stirred for 1 h at room temperature thenconcentrated under reduced pressure. The residue is neutralized with 10%aqueous K₂CO₃, extracted with EtOAc (3×20 mL), dried over Na₂SO₄,filtered and concentrated in vacuo. The crude residue is purified byPrep-HPLC to afford Example 20 (0.16 g, 33%).

The following examples are prepared in similar fashion from theappropriate reagents: Examples 21-25, 65-66, 72-74, 84-85, 93, 94, 97,99-130, 132-139, 141-143, 145, 146, and 150-153.

Synthetic Example C: Synthesis of Example 70

The mixture of R-1 (200 mg, 0.9 mmol) in DMF (3 mL) is added R-6 (333mg, 1.8 mmol). The resulting mixture is stirred at 100° C. for 2 h. Thereaction mixture is concentrated under vacuum to get a crude product(300 mg), which is dissolved in THE (5 mL) and treated with hydrazinemonohydrate (0.3 mL). The resulting mixture is stirred at 80° C. for 2h. The solvent is removed, and the residue is purified by prep-HPLC toafford Example 70 (46 mg, 25%).

Synthetic Example D: Synthesis of Example 71

To a solution of Example 10 (30 mg, 0.12 mmol) in THE (3 mL) is addedthe solution of LHMDS (1M in THF, 0.06 ml, 0.06 mmol) at −50° C.dropwise, then the mixture is stirred at −50° C. for 1 h. To thissolution is added R-7 (57 mg, 0.18 mmol) and after addition, the mixtureis stirred at 0° C. for 1 hour then adjusted to pH=6 by dilute aqueousHCl. The mixture is partitioned between water and EtOAc. The organicphase is separated, and the aqueous phase is extracted with EtOAc (12mL×3). The combined organic layer is washed with saturated aqueous NaCl,dried over Na₂SO₄, filtered, and concentrated. The residue is purifiedby Prep-HPLC to afford Example 71 (0.3 mg, 2%).

The following example was prepared in similar fashion from theappropriate reagents: Example 89.

Synthetic Example E: Synthesis of Example 78

Step 1

To a solution of R-1 (220 mg, 1.0 mmol) in DMF (4 mL) is added R-8 (210mg, 1.2 mmol) and K₂CO₃ (691 mg, 5.0 mmol), then stirred at 80° C. for 2h. The mixture is treated with H₂O (50 mL) and extracted with EtOAc (50mL×3). The combined organic layers are washed with brine (30 mL), driedover Na₂SO₄, filtered, and concentrated. The residue is purified byflash chromatography (EtOAc:Petroleum ether=1:1) to afford I-3 (310 mg,95%) [Analytical Method I, ret time=2.16 min, m/z=330.1].

Step 2

To a solution of I-3 (310 mg, 0.94 mmol) in THF (8 ml) and H₂O (2 ml) isadded LiOH.H₂O (158 mg, 3.76 mmol). The mixture is stirred at roomtemperature for 2 h. The solvent is removed under reduce pressure. Theresidue is dissolved in CH₂Cl₂/MeOH=10:1 (100 mL) and filtered. Thefiltrate is dried over Na₂SO₄ and filtered. The filtrate is concentratedunder reduce pressure to afford I-4 (280 mg, 96%) [Analytical Method E,ret time=1.87 min, m/z=316.1].

Step 3

To a solution of I-4 (280 mg, 0.88 mmol) in CH₂Cl₂ (45 mL) is addedSOCl₂ (2.09 g, 17.6 mmol). The mixture is stirred at room temperaturefor 2 h and then concentrated under reduced pressure. The residue isdissolved in THF (6 mL) then treated with a solution of ammoniumhydroxide (6 mL, 30% aqueous, 42.8 mmol) at 0° C. and stirred anadditional 0.5 h at 0° C. The reaction mixture is concentrated underreduced pressure. The residue is treated with water (10 mL) andextracted with EtOAc (20 mL×3). The combined organic layers are washedwith brine (30 mL×3), dried over Na₂SO₄, and filtered. The filtrate isconcentrated under reduce pressure then purified by prep-HPLC to affordExample 78 (130 mg, yield=48%).

The following examples are prepared in similar fashion from theappropriate reagents: Examples 79-80, 88, 90-92, 95, 96, 98, 131, 140,144, and 147-149.

Synthetic Example F: Synthesis of Example 81

The mixture of R-1 (300 mg, 1.35 mmol), R-9 (608 mg, 5.38 mmol), and2,2,6,6-tetramethylpiperidine (474 mg, 3.36 mmol) is stirred at 230° C.for 7 h. The reaction mixture is cooled to room temperature, and thenfiltered. The filtrate is directly purified by prep-HPLC to give Example78 (12.3 mg, 3%).

Synthetic Example G: Synthesis of Example 87

To a mixture of R-1 (100 mg, 0.45 mmol) and R-2 (102 mg, 0.5 mmol) in1,4-dioxane (5 mL) and H₂O (0.5 mL) is added PdCl₂(dppf) (33 mg, 0.045mmol) and K₂CO₃ (186.3 mg, 1.35 mmol). The mixture is stirred at 110° C.for 2 h under N2 the cooled, treated with H₂O (10 mL) and extracted withEtOAc (20 mL×3). The combined organic layer is washed brine (30 mL×2)and dried over Na₂SO₄. The organic layer is concentrated under reducedpressure then purified by Prep-HPLC to afford Example 87 (25 mg, 21%).

The following example is prepared in similar fashion from theappropriate reagents: Example 86.

Results are presented in Table 1:

TABLE 1 LCMS RT Mol ion Example method (min) (m/z) 1 A 1.731 330.1 2 B1.386 305.1 3 C 1.766 286 4 A 2.076 271.1 5 G 1.723 231.1 6 A 1.361348.1 8 A 1.532 296.1 9 G 1.222 319.1 10 B 1.832 257.1 11 B 1.484 273.112 E 1.678 273.0 13 C 2.128 257.0 14 G 1.892 218.1 16 B 1.539 272.1 17 H1.637 266.2 18 C 2.314 251.1 19 C 2.01 211.1 20 I 1.905 330.1 21 I 2.349315.1 22 I 2.137 275.1 23 F 1.885 270.1 24 F 2.298 255.1 25 F 2.116215.1 26 C 2.443 271 27 C 2.115 231.1 35 E 2.019 314.0 47 B 1.519 335.148 D 1.463 287.1 49 B 1.645 287.1 50 B 1.579 217.1 51 B 1.641 243.0 52 B1.626 286.0 53 A 1.918 300.1 54 E 2.206 301.0 55 F 2.042 349 56 E 1.911314.0 57 E 1.946 328.1 58 H 1.892 286.1 59 F 2.055 349 60 C 2.177 301.061 F 2.286 301.1 62 A 2.171 285.1 63 B 2.027 285.1 64 C 1.774 286.0 65 E1.628 272.0 66 B 1.467 272.1 67 B 1.411 300.0 68 B 1.455 314.1 69 E1.841 328.0 70 A 1.573 203.0 71 D 2.390 275.0 72 B 1.403 284.1 73 B1.549 284.1 74 E 1.684 298.0 75 E 2.047 287.0 76 E 2.151 273.0 77 F2.067 287.1 78 H 1.794 314.1 79 H 1.847 328.1 80 H 1.951 342.1 81 F2.574 299.1 82 G 1.945 285.2 83 B 2.013 285.1 84 B 1.484 284.1 85 G1.458 286.1 86 C 1.829 268.0 87 C 1.880 265.0 88 J 2.4 407/409 89 J 4.92290/292 90 J 3.62 308/310 91 J 4.41 385/387 92 J 4.31 290/292 93 J 3.97421 94 J 3.76 423 95 J 3.64 302/304 96 J 5.23 344/346 97 J 3.64 346/34898 J 5.08 312/314 99 J 1.96 349/351 100 J 3.96 413/415 101 J 3.67429/431 102 J 3.45 417/419 103 J 3.91 413/415 104 J 3.48 429/431 105 J3.77 413/415 106 J 3.67 399/401 107 J 2.44 421/423 108 J 2.24 385/387109 J 3.65 399/401 110 J 3.52 409/411 111 J 4.15 324/326 112 J 3.84431/433 113 J 2.35 421/423 114 J 3.56 387/389 115 J 4.3 427/429 116 J4.49 463/465 117 J 3.48 409/411 118 J 3.71 423/425 119 J 3.66 387/389120 J 3.77 423/425 121 J 3.66 387/389 122 J 3.76 423/425 123 J 3.65431/433 124 J 4.14 427/729 125 J 4.28 463/465 126 J 3.76 437/439 127 J3.11 425/427 128 J 3.31 439/441 129 J 3.25 439/441 130 J 3.05 439/441131 J 4.33 314/316 132 J 3.18 410/412 133 J 3.78 437/439 134 J 3.94451/453 135 J 3.91 451/453 136 J 2.68 425/427 137 J 3.39 444/446 138 J3.28 492/494 139 J 3.36 334/439 140 J 4.52 291 141 J 3.27 451/453 142 J3.26 399 143 J 3.27 399/401 144 J 4.44 266 145 J 1.856 360 146 J 3.72399/401 147 J 3.51 310 148 J 3.78 324 149 J 3.22 296 150 J 3.39 393/395151 J 2.19 396/398 152 J 4.14 394/396 153 J 4.13 382/384

Example 2: ARM-SAM-TIR SARM1 IC50 Assay

This Example describes an assay of ARM-SAM-TTR NADase activity and useof this assay to measure the efficacy of compounds of Formula I orFormula II to block SARM1 mediated NAD+ cleavage. This assay isoptimized in such a way as to characterize the efficacy of the compoundsin Formula I or Formula II to inhibit SARM1 activity and to calculate anIC50 value for each compound. This assay makes use of full length SARI,which encompasses the ARM, SAM and TTR domains. As demonstrated herein,expression of this fragment without the autoinhibitory N-terminal domaingenerates a constitutively active enzyme that cleaves NAD+.

Preparation of ARM-SAM-TIR Lysate (STL)

NRK1-HEK293T cells were seeded onto 150 cm² plates at 20×106 cells perplate. The next day, the cells were transfected with 15 μg ARM-SAM-TIRexpression plasmid, SEQ ID NO: 1.

(SEQ ID NO: 1)GCGATCGCGGCTCCCGACATCTTGGACCATTAGCTCCACAGGTATCTTCTTCCCTCTAGTGGTCATAACAGCAGCTTCAGCTACCTCTCAATTCAAAAAACCCCTCAAGACCCGTTTAGAGGCCCCAAGGGGTTATGCTATCAATCGTTGCGTTACACACACAAAAAACCAACACACATCCATCTTCGATGGATAGCGATTTTATTATCTAACTGCTGATCGAGTGTAGCCAGATCTAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGCTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTGGTTTAGTGAACCGTCAGATCAGATCTTTGTCGATCCTACCATCCACTCGACACACCCGCCAGCGGCCGCTGCCAAGCTTCCGAGCTCTCGAATTCAAAGGAGGTACCCACcatgGCCATGCATCACCACCACCATCATAGCTCCGGCGTCGACCTCGGCACCGAGAATTTATATTTCCAAAGCGGCCTCAATGATATCTTCGAGGCCCAGAAGATCGAGTGGCACGAGGGCAGCTCCGACCTCGCCGTGCCCGGTCCCGATGGAGGCGGAGGCACTGGTCCTTGGTGGGCTGCTGGCGGCAGAGGCCCTAGAGAAGTGAGCCCCGGTGCTGGCACCGAGGTGCAAGACGCTCTGGAGAGGGCTCTGCCCGAACTGCAGCAAGCTCTGTCCGCTTTAAAGCAAGCTGGAGGAGCTAGAGCCGTCGGCGCCGGACTGGCCGAAGTGTTCCAGCTCGTGGAGGAAGCTTGGTTATTACCCGCTGTGGGAAGAGAGGTCGCCCAAGGTCTGTGTGACGCCATTCGTCTGGACGGAGGTTTAGACTTATTACTGAGGCTGCTGCAAGCTCCCGAACTGGAGACAAGGGTCCAAGCTGCTCGTCTGCTGGAGCAGATCCTCGTGGCCGAGAATCGTGACAGAGTGGCTAGAATCGGTTTAGGCGTCATCCTCAATTTAGCCAAAGAGAGGGAGCCCGTTGAGCTGGCCAGAAGCGTCGCTGGCATCCTCGAGCACATGTTCAAGCATTCCGAGGAGACTTGTCAGAGACTGGTCGCCGCCGGAGGACTCGATGCTGTTTTATACTGGTGCAGAAGGACAGACCCCGCTTTACTGAGGCATTGTGCTCTGGCCCTCGGCAATTGCGCTTTACATGGAGGCCAAGCCGTCCAGAGAAGGATGGTGGAGAAAAGAGCCGCCGAGTGGCTGTTCCCTTTAGCCTTCTCCAAAGAAGACGAACTGTTACGTCTGCATGCTTGTCTCGCTGTCGCTGTTTTAGCCACCAACAAGGAGGTGGAAAGGGAAGTGGAAAGAAGCGGAACACTGGCTTTAGTCGAACCTCTGGTGGCTTCTTTAGATCCCGGAAGGTTTGCCAGATGTCTGGTCGACGCCAGCGATACCTCCCAAGGAAGAGGCCCCGACGATCTCCAGAGACTGGTGCCTCTGCTGGACAGCAATCGTCTGGAGGCCCAATGTATTGGCGCCTTCTATCTCTGCGCCGAAGCCGCCATCAAGTCTTTACAAGGTAAGACCAAGGTGTTCTCCGACATTGGAGCCATCCAATCTTTAAAGAGGCTGGTGAGCTATTCCACCAACGGCACAAAAAGCGCTTTAGCCAAAAGAGCTTTAAGACTGCTGGGCGAAGAGGTGCCTAGGCCCATTTTACCTTCCGTGCCTAGCTGGAAGGAGGCCGAGGTGCAGACTTGGCTGCAGCAGATCGGCTTTAGCAAATATTGCGAATCCTTTAGGGAGCAGCAAGTTGACGGCGATTTATTATTAAGGCTGACCGAGGAAGAGCTCCAGACAGATTTAGGCATGAAAAGCGGCATCACTCGTAAGAGGTTCTTTCGTGAGCTCACCGAACTGAAGACCTTCGCCAACTACTCCACTTGTGATCGTAGCAATTTAGCTGATTGGCTCGGATCCCTCGATCCCAGATTTCGTCAGTACACCTATGGACTCGTCTCTTGTGGACTGGACAGATCTTTACTGCATCGTGTGAGCGAGCAACAGCTGCTGGAAGATTGCGGCATCCATTTAGGAGTGCACAGAGCCAGAATTCTGACCGCCGCTAGAGAGATGCTGCATTCCCCTCTCCCTTGTACCGGAGGCAAGCCTAGCGGAGACACCCCCGACGTGTTCATCAGCTATCGTAGAAACAGCGGAAGCCAGCTGGCCTCTTTACTGAAGGTCCATTTACAGCTGCACGGATTTAGCGTCTTCATCGACGTGGAGAAACTGGAGGCTGGCAAGTTCGAGGACAAGCTGATCCAGTCCGTGATGGGCGCTAGGAATTTCGTTTTAGTGCTCAGCCCCGGCGCTCTGGATAAATGCATGCAAGATCATGACTGTAAGGACTGGGTCCACAAGGAAATCGTGACCGCTCTGTCTTGTGGCAAGAACATCGTCCCCATCATCGACGGCTTCGAATGGCCCGAGCCTCAAGTTCTCCCCGAAGATATGCAAGCTGTTTTAACCTTCAATGGAATCAAGTGGAGCCACGAGTACCAAGAAGCCACAATCGAGAAGATCATTCGTTTTCTGCAAGGTAGATCCTCCAGAGATTCCTCCGCTGGCAGCGACACATCTTTAGAGGGCGCCGCCCCTATGGGTCCTACCTAATAATctagAAGTTGTCTCCTCCTGCACTGACTGACTGATACAATCGATTTCTGGATCCGCAGGCCTCTGCTAGCTTGACTGACTGAGATACAGCGTACCTTCAGCTCACAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGGTGTGGGAGGTTTTTTAAAGCAAGTAAAACCTCTACAAATGTGGTATTGGCCCATCTCTATCGGTATCGTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGTGCCCCTCGGGCCGGATTGCTATCTACCGGCATTGGCGCAGAAAAAAATGCCTGATGCGACGCTGCGCGTCTTATACTCCCACATATGCCAGATTCAGCAACGGATACGGCTTCCCCAACTTGCCCACTTCCATACGTGTCCTCCTTACCAGAAATTTATCCTTAAGGTCGTCAGCTATCCTGCAGGCGATCTCTCGATTTCGATCAAGACATTCCTTTAATGGTCTTTTCTGGACACCACTAGGGGTCAGAAGTAGTTCATCAAACTTTCTTCCCTCCCTAATCTCATTGGTTACCTTGGGCTATCGAAACTTAATTAACCAGTCAAGTCAGCTACTTGGCGAGATCGACTTGTCTGGGTTTCGACTACGCTCAGAATTGCGTCAGTCAAGTTCGATCTGGTCCTTGCTATTGCACCCGTTCTCCGATTACGAGTTTCATTTAAATCATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCATTTAAATTTCCGAACTCTCCAAGGCCCTCGTCGGAAAATCTTCAAACCTTTCGTCCGATCCATCTTGCAGGCTACCTCTCGAACGAACTATCGCAAGTCTCTTGGCCGGCCTTGCGCCTTGGCTATTGCTTGGCAGCGCCTATCGCCAGGTATTACTCCAATCCCGAATATCCGAGATCGGGATCACCCGAGAGAAGTTCAACCTACATCCTCAATCCCGATCTATCCGAGATCCGAGGAATATCGAAATCGGGGCGCGCCTGGTGTACCGAGAACGATCCTCTCAGTGCGAGTCTCGACGATCCATATCGTTGCTTGGCAGTCAGCCAGTCGGAATCCAGCTTGGGACCCAGGAAGTCCAATCGTCAGATATTGTACTCAAGCCTGGTCACGGCAGCGTACCGATCTGTTTAAACCTAGATATTGATAGTCTGATCGGTCAACGTATAATCGAGTCCTAGCTTTTGCAAACATCTATCAAGAGACAGGATCAGCAGGAGGCTTTCGCATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCGCGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGCTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTATTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATTGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACCTTGCGTAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAGTTGATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACCGATTCTAGGTGCATTGGCGCAGAAAAAAATGCCTGATGCGACGCTGCGCGTCTTATACTCCCACATATGCCAGATTCAGCAACGGATACGGCTTCCCCAACTTGCCCACTTCCATACGTGTCCTCCTTACCAGAAATTTATCCTTAAGATCCCGAATCGTTTAAACTCGACTCTGGCTCTATCGAATCTCCGTCGTTTCGAGCTTACGCGAACAGCCGTGGCGCTCATTTGCTCGTCGGGCATCGAATCTCGTCAGCTATCGTCAGCTTACCTTTTTGGCA.

The cultures were supplemented with 1 mM NR at time of transfection tominimize toxicity from ARM-SAM-TIR overexpression. Forty-eight hoursafter transfection, cells were harvested, pelleted by centrifugation at1,000 rpm (Sorvall ST 16R centrifuge, Thermo Fisher), and washed oncewith cold PBS (0.01 M phosphate buffered saline NaCl 0.138 M; KCl 0.0027M; pH 7.4). The cells were resuspended in PBS with protease inhibitors(cOmplete™ protease inhibitor cocktail, Roche product #11873580001) andcell lysates were prepared by sonication (Branson Sonifer 450, output=3,20 episodes of stroke). The lysates were centrifuged (12,000×g for 10min at 4° C.) to remove cell debris and the supernatants (containingARM-SAM-TIR protein) were stored at −80° C. for later use in the invitro ARM-SAM-TIR NADase assay (see below). Protein concentration wasdetermined by the Bicinchoninic (BCA) method and used to normalizelysate concentrations.

ARM-SAM-TIR IC50 Assay of Formula I or Formula II Compounds.

The enzymatic assay was performed in a 384-well polypropylene plate inDulbecco's PBS buffer in a final assay volume of 20 μL. ARM-SAM-TIRlysate with a final concentration of 5 μg/mL was pre-incubated with therespective compound at 1% DMSO final assay concentration over 2 h atroom temperature. The reaction was initiated by addition of 5 μM finalassay concentration of NAD+ as substrate. After a 2 hr room temperatureincubation, the reaction was terminated with 40 μL of stop solution of7.5% trichloroactetic acid in acetonitrile. The NAD+ and ADPRconcentrations were analyzed by a RapidFire High Throughput MassSpectrometry System (Agilent Technologies, Santa Clara, Calif.) using anAPI4000 triple quadrupole mass spectrometer (AB Sciex Framingham,Mass.).

Results are presented below in Table 2. Compounds having an activitydesignated as “A” provided an IC₅₀<5 μM; compounds having an activitydesignated as “B” provided an IC₅₀ 5-15 μM; compounds having an activitydesignated as “C” provided an IC₅₀ 15.01-30 μM; compounds having anactivity designated as “D” provided an IC₅₀>30 μM; nd: not determined.

TABLE 2 SARM1 Example IC₅₀ (μM) 1 B 2 D 3 B 4 A 5 A 6 D 7 nd 8 D 9 D 10A 11 A 12 A 13 A 14 A 15 D 16 D 17 B 18 B 19 A 20 A 21 A 22 A 23 D 24 A25 B 26 A 27 A 28 D 29 C 30 B 31 A 32 D 33 B 34 B 35 A 36 A 37 A 38 C 39D 40 A 41 C 42 A 43 C 44 A 45 A 46 A 47 A 48 A 49 A 50 A 51 D 52 D 53 D54 A 55 B 56 B 57 A 58 A 59 A 60 A 61 A 62 A 63 A 64 A 65 A 66 A 67 A 68B 69 A 70 C 71 A 72 A 73 A 74 A 75 A 76 A 77 A 78 A 79 A 80 A 81 A 82 A83 A 84 B 85 B 86 B 87 C 88 A 89 A 90 A 91 A 92 A 93 A 94 A 95 A 96 A 97A 98 A 99 A 100 A 101 A 102 A 103 A 104 A 105 B 106 A 107 A 108 A 109 A110 A 111 A 112 A 113 A 114 A 115 A 116 A 117 A 118 A 119 A 120 A 121 A122 A 123 A 124 A 125 A 126 A 127 A 128 A 129 A 130 A 131 A 132 A 133 A134 A 135 A 136 A 137 A 138 A 139 A 140 A 141 A 142 A 143 A 144 A 145 A146 A 147 A 148 A 149 A 150 A 151 A 152 A 153 A

1. A compound of Formula I-c:

or a pharmaceutically acceptable salt thereof, wherein: R¹ is selectedfrom the group consisting of hydrogen, halogen, N(R)₂, OR, C₁₋₆aliphatic, a 3- to 7-membered monocyclic heterocyclic ring having 1-2heteroatoms independently selected from nitrogen, oxygen, and sulfur, a5- to 6-membered heteroaryl ring having 1-3 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur, and a 7- to 8-memberedbridged bicyclic heterocyclic ring having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur, wherein each of C₁₋₆aliphatic, monocyclic or bridged bicyclic heterocyclic ring, andheteroaryl ring is optionally substituted with 1-3 R¹* groups; R¹* isselected from oxo, halogen, N(R)₂, OR, C(O)R, S(O)₂R, C(O)OR, C(O)N(R)₂,optionally substituted C₁₋₆ aliphatic, and an optionally substituted 5-to 6-membered heteroaryl ring having 1-3 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur; R is selected from the groupconsisting of hydrogen, an optionally substituted C₁₋₆ aliphatic, and anoptionally substituted 3- to 7-membered saturated monocyclic ring having0-2 heteroatoms independently selected from nitrogen, oxygen, andsulfur, or: two R groups, together with the nitrogen atom to which theyare attached, form an optionally substituted 3- to 6-memberedheterocyclic ring containing 0-1 additional heteroatom selected fromnitrogen, oxygen, or sulfur; R² is selected from the group consisting ofhydrogen, halogen, S(O)₂R, C(O)OR, C(O)N(R)₂, and an optionallysubstituted group selected from C₁₋₆ aliphatic, a 3- to 7-memberedheterocyclic ring having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, and a 5- to 6-membered heteroaryl ringhaving 1-3 heteroatoms independently selected from nitrogen, oxygen, andsulfur; and R³ is selected from the group consisting of hydrogen,halogen, CN, CF₃, C₁₋₃ aliphatic, and —C(O)N(R^(a)*)₂. 2.-6. (canceled)7. The compound according to claim 1, wherein R¹ is selected from


8. The compound according to claim 1, wherein R¹ is N(R)₂.
 9. Thecompound according to claim 8, wherein each R is hydrogen.
 10. Thecompound according to claim 8, wherein each R is —CH₃.
 11. The compoundaccording to claim 8, wherein R¹ is —NHCH₃.
 12. The compound accordingto claim 8, wherein R¹ is


13. The compound according to claim 8, wherein R¹ is


14. The compound according to claim 1, wherein R¹ is —OR.
 15. Thecompound according to claim 14, wherein R is hydrogen.
 16. The compoundaccording to claim 14, wherein R is —CH₃.
 17. The compound according toclaim 1, wherein R² is halogen.
 18. The compound according to claim 17,wherein R² is chloro.
 19. The compound according to claim 17, wherein R²is bromo.
 20. The compound according to claim 17, wherein R² is fluoro.21. The compound according to claim 1, wherein R² is —CH₃.
 22. Thecompound according to claim 1, wherein R² is hydrogen.
 23. The compoundaccording to claim 1, wherein R³ is hydrogen.
 24. The compound accordingto claim 1, wherein R³ is halogen.
 25. The compound according to claim24, wherein R³ is chloro.